Bis-14-membered ring diketal diamines: Synthesis and evaluation of their anti-HIV and anti-tumoral activities

Article abstract of DOI:10.1016/j.ejmech.2009.03.008

Chiral and achiral macrocyclic bis-diketal diamines, analogs of bicyclam AMD 3100, were synthesized in three steps from the previously obtained 14-membered ring diketal dilactams. Their monoreduction with lithium aluminium hydride gave the corresponding d

Full text of DOI:10.1016/j.ejmech.2009.03.008

European Journal of Medicinal Chemistry 44 (2009) 3138–3146
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Bis-14-membered ring diketal diamines: Synthesis and evaluation of their
anti-HIV and anti-tumoral activities
a
a
b
a,
*
´
Radouane Affani , Franck Pelissier , Anne-Marie Aubertin , Denise Dugat
a
`
`
`
´ ˆ
´
Laboratoire SEESIB (Synthese Et Etude de Systemes a Interet Biologique), UMR CNRS 6504, Universite Blaise Pascal de Clermont-Ferrand, 24 Avenue des Landais,
`
63177 Aubiere, France
b Institut de Virologie, INSERM U778, Universite´ Louis Pasteur, 67000 Strasbourg, France
a r t i c l e i n f o
a b s t r a c t
Article history:
Chiral and achiral macrocyclic bis-diketal diamines, analogs of bicyclam AMD 3100, were synthesized in
three steps from the previously obtained 14-membered ring diketal dilactams. Their monoreduction with
lithium aluminium hydride gave the corresponding diketal aminolactams. Coupling these with dibromo-
p-xylene led to xylyl dimer compounds. A second reduction step yielded the expected bis-diketal
diamines in the methyl and unsubstituted series. Biological tests on the unreduced and reduced dimers
showed both weak anti-HIV and anti-proliferative activities for the bis-diphenyl diketal aminolactam
13b, with a mode of action probably different from that of AMD 3100.
Received 24 July 2008
Received in revised form
24 February 2009
Accepted 12 March 2009
Available online 24 March 2009
Keywords:
Bis-diketal diamine macrocycles
AMD 3100 analogs
Ó 2009 Elsevier Masson SAS. All rights reserved.
Anti-viral and anti-tumoral activities
CXCR4 antagonists
1. Introduction
In continuation of our work on the synthesis, conformational
analysis and binding properties of new macrocyclic 14-membered
Macrocyclic saturated polyamines have generated continuous
interest because of their physicochemical and biological properties
[1]. Among these derivatives, bicyclams form a new class of
compounds that exhibit anti-viral [1b,2–7] and anti-tumoral [7–9]
activities as antagonists of the chemokine co-receptor CXCR4 of the
cell membrane [3–9]. As anti-HIV-1 agents, they interfere at the
virus-cell fusion step in blocking the interaction between CXCR4
and the glycoproteins gp 120 of the viral envelope, i.e. the entry of
the virus into the host cell, and thereby its replication [3–7]. As
anti-tumoral agents, they block the binding of CXCR4 with its
ring diketal dilactams [11] and diketal diamines [12], we report the
synthesis of substituted bis-dioxygenated analogs of bicyclam AMD
3100 (Fig. 1) and the evaluation of their anti-viral and anti-tumoral
activities.
2. Chemistry
2.1. Synthesis
Bis-diketal diamines were targeted by a coupling reaction of
diketal aminolactams (Fig. 1), previously obtained as minor prod-
ucts in the synthesis of diketal diamines by reduction of the cor-
responding diketal dilactams [12].
natural ligand, the chemokine C-X-C, SDF-1a (stromal cell-derived
factor), and hence inhibit growth, migration and proliferation of
cancerous cells [7–9]. One of these compounds, the xylyl bicyclam
AMD 3100, several synthesis of which have been reported
[2b,3c,10], has so far proved to be the most specific CXCR4 antag-
onist with a nanomolar binding affinity [2a,3a,4c,5–7]. However,
this molecule presents poor oral bioavailability due to its highly
polar nature [5,6], and possible cardiac side effects disclosed after
Phase II trials [6]. In this light, novel chemical analogs of this
bioactive agent were likely to be of interest.
2.1.1. Synthesis of diketal aminolactams
The study was conducted on the two major isomers b (trans-
OMe) and c (cis-OMe) [13] of the three already developed series of
diketal dilactams (1: R1 ¼ Ph, R2 ¼ H; 2: R1 ¼ H, R2 ¼ Me; 3:
R1 ¼ R2 ¼ H), using lithium aluminium hydride (10 equiv.) as
reducing agent [12]. However, some modifications were made to
the previously described reaction conditions to invert the ratio of
singly reduced compounds 4–6 to doubly reduced compounds 7–9
(Fig. 2, Table 1). As previously, small amounts of NEt3 were added to
* Corresponding author. Tel.: þ33 04 73 40 71 27; fax: þ33 04 73 40 77 17.
E-mail address: denise.dugat@univ-bpclermont.fr (D. Dugat).
0223-5234/$ – see front matter Ó 2009 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2009.03.008

R. Affani et al. / European Journal of Medicinal Chemistry 44 (2009) 3138–3146
3139
R2
Ph
Ph
3
R1
OMe
OMe
Y'
2
Y
Y
OMe
Y'
Y
NH
O
NH O14
NH HN
NH HN
NH
O
O
6
13
7
O
HN
O
HN
HN
9
Y'
MeO
MeO
MeO
10 Ph
R1
Ph
R2
Cyclam
1b
Y = Y' = O
1c
4c
Diketal dilactams
Diketal diamines
Diketal aminolactams
: Y = Y' = O
4b
7b
Y = O ; Y' = H2
Y = Y' = H2
: Y = Y' = H2
: Y = O; Y' = H2
7c
Me
Me
NH HN
NH
OMe
Y'
OMe
Y
Y
NH
O
NH
O
N
HN
N
Y
O
HN
NH HN
O
HN
Y'
MeO
Bicyclams
MeO
Me
Me
AMD 3100 : Y =
Y = Y' = O
2c
5c
2b
R2
5b
8b
Y = O ; Y' = H2
Y = Y' = H2
R1
R2
R1
8c
OMe
Y
NH
O
N
OMe
O
N
OMe
OMe
Y'
Y
Y
O
NH
O
O
NH
O
O
MeO
O
HN
R1
Y
MeO
R2
HN
R1
HN
R2
Bis-diketal aminolactams : Y = O
Bis-diketal diamines : Y = H2
Y'
Y = Y' = O
MeO
MeO
3c
6c
3b
6b
9b
Y = O ; Y' = H2
Y = Y' = H2
9c
Fig. 1. General structure of cyclam, diketal dilactams, diketal diamines, diketal ami-
nolactams, bicyclams, bis-diketal dilactams, and bis-diketal diamines.
R2
R1
OMe
O
10 : R1 = Ph ; R2 = H ; 14α-OMe
11 : R1 = H ; R2 = Me ; 14β-OMe
12 : R1 = R2 = H ; 14β-OMe
minimize the effect of an elimination side reaction leading to
unsaturated ketal aminolactams 10–12 [12].
NH O14
HN
6
Comparison of entries 1 and 2 (Table 1), which recall two
earlier experiments carried out from phenyl diketal dilactam 1c
[12a], showed that a lower substrate concentration favoured the
formation of diketal aminolactam 4c, the yield of which could be
optimized in the conditions of entry 3. Entry 4 reports the
optimal formation conditions of both diketal aminolactam 4b and
diketal diamine 7b, the reaction always leading mainly to the
mono-reduced compound independently of the concentration
[12b]. In the methyl series, a better yield of aminolactam 5b was
obtained in the conditions of entry 6 (c ¼ 8 ꢀ 10ꢁ3 M) compared
with the previously reported conditions of entry 5 [12b]. In
contrast, the formation of 5c was only perceptible (yield 12%,
conversion rate 60%) on decreasing the excess of LAH (6 equiv.)
and the reaction time (9 h) (entry 9 compared with entries 7 and
8). In the unsubstituted series the best formation conditions of
diketal aminolactams 6b and 6c were found at concentrations
5 ꢀ10ꢁ3 M and 3 ꢀ10ꢁ3 M respectively, using 10 equiv. of LAH
(entries 10 and 11).
O
7
R1
R2
Fig. 2. Structure and stereochemistry of compounds 1b,c-9b,c, 10–12.
resulting from complexation between the diketal diamines gener-
ated and the Liþ cation, which could therefore no longer catalyse
the reaction [14].
2.1.2. Synthesis of bis-diketal aminolactams
The coupling step between diketal aminolactams 4–6 and the
a,a
0-dibromo-p-xylene was studied from the substrates obtained in
appreciable yields, i.e. from 4b,c, 5b, 6b,c. The reactions were per-
formed with minor variations of literature procedures [2b,3c,10],
using K2CO3 as basic agent [2b,3c]. They led to the expected bis-
diketal aminolactams 13–15, obtained in moderate to good yields
(37–72%), and accompanied in both substituted series by small
amounts (6–19%) of bromo ketal aminolactams 16–17, which
correspond to single coupling products (Scheme 1).
As previously reported [12], in all the series the reduction
occurred with moderate total yields (43–53% for the selected
entries) that were difficult to improve. These results may be
explained by the presence of polar compounds (Table 1) probably

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R. Affani et al. / European Journal of Medicinal Chemistry 44 (2009) 3138–3146
Table 1
namely the addition of a commercial THF hydride solution to the
substrate solution, was adopted due to the small amounts of
starting materials available.
Reduction of diketal dilactams 1b,c, 2b,c and 3b,c by LAH (10 equiv) in THF, and in
the presence of small amounts of NEt3 (0.1–0.2 equiv.).
Entry
Substrate
(R)
csubstrate
Time
(h)
Conv.
(%)d
Resultse
(%)
In the phenyl series, the reduction performed at different
substrate concentrations and reaction times led to either an incom-
plete reaction giving rise to a mixture of singly and doubly reduced
bis-macrocycles difficult to separate (csubstrate ¼ 30 ꢀ 10ꢁ3 M, 10 h
and 40 ꢀ 10ꢁ3 M, 9 h), or a major cleavage reaction regenerating
diphenyl diketal diamine 7b (csubstrate ¼ 50 and 60 ꢀ 10ꢁ3 M, 8 h).
In contrast, in the methyl and unsaturated series, diketal
diamines 18b and 19c were obtained with 26% and 22% yields
respectively (conversion rate 100%) with the following substrate
concentrations and reaction times: 18b, csubstrate ¼ 100 ꢀ 10ꢁ3 M,
9 h; 19c, csubstrate ¼ 60 ꢀ 10ꢁ3 M, 6 h (Scheme 2). In both cases, the
reduction was also partially accompanied by a benzyl cleavage
giving rise to the corresponding diketal diamines 8b and 9c, in low
yield (9%) compared with the phenyl series.
In summary, the access to bis-diketal diamines succeeded in the
methyl and unsubstituted series but failed in the phenyl series. The
difficulties observed in the reduction of the bis-phenyl amino-
lactam 13b, in which the 3,30-Ph, and 7,70-OMe groups are in a trans
position, are probably due to the hindrance of the two faces of the
molecule at the level of the 5,50-CO carbonyl groups, as previously
reported in the reduction of trans-OMe phenyl diketal dilactam 1b
[12b]. In the two successful series, the reduction took place again
with moderate total yields (30–35%) which were difficult to
improve as in the formation of compounds 4–9 above.
(10ꢁ3 M)
1a
2b
3
4
5
1c (Ph)
1c (Ph)
1c (Ph)
1b (Ph)
2b (Me)
2b (Me)
2c (Me)
2c (Me)
2c (Me)
3b (H)
17
25
8
14
12
8
10
5
4
9
11
9
10
18
18
20
20
9
90
95
84
85
93
88
85
85
60
78
70
4c (16), 7c (21), 10 (14)
4c (8), 7c (30), 10 (8)
4c (35), 7c (10), 10 (8)
4b (48), 7b (4)
5b (21), 8b (20), 11 (2)
5b (28), 8b (12), 11 (2)
8c (17)
6
7
8
5c (1), 8c (12)
9c
10
11
5c (12), 8c (22), 11 (1)
6b (27), 9b (16), 12 (2)
6c (29), 9c (14)
5
3
9
10
3c (H)
a
Absence of NEt3.
b
c
Molar ratio of NEt3/substrate ¼ 0.4/1.
Molar ratio of LAH/substrate ¼ 6:1.
d
e
Conversion rate based on the amount of recovered starting material.
Isolated yield by column chromatography; at the end of the process, unidenti-
fied polar compounds were invariably isolated in 7–22% yield, which increased with
the reaction time.
2.1.3. Synthesis of bis-diketal diamines
Due to the moderate yields of 13c and 15b, the reduction of bis-
diketal aminolactams was studied on only 13b, 14b and 15c, i.e. on
the isomer of each series obtained with the best yield. The reactions
were repeated with LAH (10 equiv.) in THF and in the presence of
NEt3 (0.02 equiv./LAH). An inverse addition order of the reactants,
R2
R2
R1
R2
R2
2
OMe
R1
R1
R1
O
2
OMe
9
OMe
O
OMe
15
NH O14
O
NH O14
NH O14
6
N
O
7
7
N12
R1
6
+
Br
7
O
6
7
13
10
O
N
O
HN
R1
14 O HN
9
MeO
15
MeO
MeO
9
3
O
MeO
R1
2
R2
R1
R2
R2
R2
R1 = Ph ; R2 = H ; 7β-OMe ; 14α-OMe
R1 = Ph ; R2 = H ; 7α-OMe ; 14α-OMe
R1 = H ; R2 = Me ; 7α-OMe ; 14β-OMe
4b
13b (61%)
13c (37%)
14b (72%)
16b (16%)
16c (19%)
4c
5b
17b (6%)
R1 = R2 = H ; 7α-OMe ; 14β-OMe
R1 = R2 = H ; 7β-OMe ; 14β-OMe
-
-
6b
6c
15b (48%)
15c (55%)
Scheme 1. Reagents and conditions:
a,
a
0-dibromo-p-xylene (0.5–0.6 equiv.), K2CO3 (1.0–1.2 equiv.), CH3CN (csubstrate ¼ 50 ꢀ 10ꢁ3 M),
D
, 8–10 h.
R2
R2
R1
R1
R2
R2
OMe
OMe
R1
R1
O
OMe
O
OMe
NH O14
NH O 14
N
O 7
HN
R1
N
O
7
7
7
O
N
O
N
14
14 O HN
MeO
O
MeO
R1
R1
MeO
MeO
R2
R2
R1
R2
R2
R1 = H ; R2 = Me ; 7α-OMe
R1 = R2 = H ; 7β-OMe
18b (26%)
19c (22%)
14b
15c
Scheme 2. Reagents and conditions: LAH (10 equiv.), NEt3 (0.2 equiv.), THF (csubstrate ¼ 60–100 ꢀ 10ꢁ3 M),
D, 6–9 h.

R. Affani et al. / European Journal of Medicinal Chemistry 44 (2009) 3138–3146
3141
Table 2
CXCR4 receptor to enter their target cells [7b], the effects of the
above chemical entities were studied in vitro with two T cell lines,
MT-4 and CEM-SS, infected by X4 viral isolates. The activity of
a given compound on the replication of HIV-1 was measured by
quantification of (i) the inhibition of virus-induced cytopathoge-
nicity in infected MT-4 cells, and (ii) the reverse transcriptase (RT)
activity associated with virus particles released from infected
CEM-SS cells.
The 50% cytotoxic concentration (CC50) was evaluated in parallel
to the 50% inhibitory concentration (IC50) (Table 2). Among the
tested compounds, only the bis-diphenyl diketal aminolactam 13b
showed an inhibitory activity in regard to HIV-1 infection in
CEM-SS cells, but with lower values than AMD 3100.
Anti-viral activity and cytotoxicity data of bis-diketal aminolactams 13b,c, 15b, and
bis-diketal diamines 18b, 19c, evaluated on MT-4 cells and on CEM-SS cells infected
with X4 HIV strains (HIV-1IIIB and HIV-1 LAI respectively).a
Compound (R)
HIV-1 IIIB/MT4
HIV-1 LAI/CEM-SS
b
c
b
c
IC50
(m
M)
CC50
(
m
M)
IC50
(
m
M)
CC50 (mM)
13b (Ph)
13c (Ph)
15b (H)
>CC50
>10
>100
>100
> CC50
0.41
7.8
>10
8.65
>10
>10
>10
>100
>100
50
>100
>100
>50
0.32
>100
>100
>50
>5
18b (Me)
19c (H)
AMD 3100d
6.5
a
All data represent mean values of 2 separate experiments.
IC50: 50% inhibitory concentration ¼ concentration needed to inhibit 50% virus
b
replication in vitro.
c
CC50: 50% cytotoxic concentration ¼ concentration required to cause 50% death
3.2. Anti-tumoral activity evaluation [15]
of uninfected cells.
d
Commercial sample purchased from CheMatech.
The in vitro anti-proliferative activities of compounds 13b,c,
15b, 18b and 19c were evaluated in triplicate towards ten human
cell lines [buccal carcinoma (KB), glioblastoma (SF268), myeloid
leukaemia (HL60), mammary carcinoma (MCF7 and MCF7R),
colon carcinoma (HCT116), pulmonary carcinoma (A549), pros-
tate adenocarcinoma (PC3), ovary adenocarcinoma (OV3) and
hepatocarcinoma (HepG2)] using the classic colorimetric MTS
test (Table 3). Taxotere was used as a positive control at a drug
concentration of 2.5 ꢀ10ꢁ10 M. The percentage of cell prolifera-
tion inhibition was defined as absorbance in experimental wells
compared with absorbance in control wells, after subtraction of
the blank values. Under the conditions used (10ꢁ5 M final
concentration of tested compounds), the bis-diphenyl diketal
aminolactam 13b showed a mild cytotoxic activity of about 50%
towards five cell lines (KB, HL60, MCF7, HCT116 and PC3), i.e.
2.2. Characterization
The structure and stereochemistry of all the derivatives 4–19
were established from IR, MS, 1D NMR [1H, 13C (J MOD)] and 2D
NMR [COSY 1H-1H, HSQC (heteronuclear single quantum correla-
tion) and sometimes HMBC (heteronuclear multiple bond correla-
tion)] spectroscopic data, which allowed identification of all the
hydrogen and carbon atoms.
The characteristics of the monomer compounds 4–12 have
already been reported [12b]. Briefly, (i) in the 1H and 13C NMR
spectra, diamines 7c, 8c, 9b,c, which all possess a symmetry
element (C2 for 7c, 8c, 9c, center for 9b), show only one signal for
each pair of identical groups of the macrocycle, whereas diamines
7b and 8b exhibit double signals due to the asymmetry of the two
ring chains, and (ii) the reduced carbonyl of trans-OMe amino-
lactams 4b and 5b correspond to the 12-CO carbonyl of chain 2 in
which the 14-OMe group and the 3,10-R substituents are in a cis
position [12b].
a
cytotoxicity corresponding approximately to an IC50 of
z10 M.
m
4. Discussion
The 1H and 13C NMR spectra of the dimer compounds 13–19 are
also simplified due to the presence of a C2 axis perpendicular to the
aromatic linker.
Biological tests, performed on the synthesized bis-macrocyclic
compounds 13b,c, 15b, 18b, 19c gave an encouraging result for
one analog compared with the others: bis-phenyl diketal ami-
nolactam 13b showed both a weak anti-HIV-1 activity and some
anti-proliferative effects. The interaction of AMD 3100 with the
receptor CXCR4 has been described as probably due to binding of
two antiperiplanar NH amine functions of the two cyclam moie-
ties with the free carboxylic acid groups of Asp171 and Asp262
located on respectively transmembrane TM-IV and TM-VI of the
receptor [4b,6b,7]. This is not possible in 13b since the two NH
concerned are involved in amide links. Hence the activity of this
3. Biological assays
Anti-HIV and anti-tumoral tests were performed on the two
synthesized bis-diketal diamines 18b, 19c, and on the three bis-
diketal aminolactams 13b,c, 15b, not involved in the last reduction
step.
3.1. Anti-HIV activity evaluation
compound probably depends on other factors e.g. p–p stacking
effects between the 3,30,10,100-phenyl substituents of the mole-
cule and aromatic rings of Phe, Tyr and (or) Trp units of the
transmembrane.
Since the anti-HIV activity of AMD 3100 appears to be confined
to the T-lymphotropic (X4) HIV strains, i.e. those strains that use the
Table 3
Percentage cell proliferation inhibition of bis-diketal aminolactams 13b,c, 15b, and bis-diketal diamines 18b, 19c, at a drug concentration of 10ꢁ5 M.a
Compound
KB
SF268
HL60
MCF7
MCF7R
HCT116
A549
PC3
OV3
HepG2
13bb
13c
15b
47
12
00
11
00
75
27
14
00
05
12
37
54
31
10
08
08
19
45
25
13
00
08
0
14
08
02
04
04
0.03
47
21
07
10
11
24
33
17
00
00
00
14
51
15
00
09
10
9
38
18
00
08
15
22
27
00
04
00
00
0
18b
19c
Taxoterec
a
All experiments were carried out in triplicate with DMSO as solvent.
b
c
For 13b, the best observed values (z50%) correspond approximately to an IC50 z 10 mM.
Data from Ref. [15].Taxotere was used as a positive control at a drug concentration of 2.5 ꢀ10ꢁ10 M.

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R. Affani et al. / European Journal of Medicinal Chemistry 44 (2009) 3138–3146
5. Conclusion
chromatography (silicagel, Et2O/MeOH) yielded diketal amino-
lactams and diketal diamines.
Variously substituted bis-diketal aminolactams and bis-diketal
diamines were prepared in two and three steps respectively from
previously and rapidly synthesized macrocyclic diketal dilactams
[11a]. The first step, which corresponded to the formation of singly
6.1.2.1. (3R,7R,10R,14S)-Diphenyl
(3R,7R,10R,14S)-diphenyl diketal diamine (7b). The preparation of
these compounds was described previously [12b].
diketal
aminolactam
(4b),
reduced compounds 4–6, was efficient in
a
3–14 ꢀ10ꢁ3 M
concentration range for five of the six series, but failed from the cis
methyl dilactam 2c, the reduction of which led mainly to diamine
8c, irrespective of the concentration. After a coupling stage which
gave bis-diketal aminolactams 13–15 in 37–72% yield, the final
reduction step allowed the isolation of bis-diketal diamines 18b
(R ¼ Me) and 19c (R ¼ H), while a partial reaction or a major benzyl
cleavage accompanied the reduction of the phenyl product 13b.
Biological assays made on the unreduced and reduced dimer
macrocycles showed that the bis-diphenyl diketal aminolactam 13b
was the most promising compound, with both weak anti-HIV and
anti-proliferative activities. These results lead to consider a mode of
action different from that of AMD 3100, and indicate the probable
6.1.2.2. (3R,7S,10R,14S)-Diphenyl
diketal
aminolactam
(4c),
(3R,7S,10R,14S)-Diphenyl diketal diamine (7c), and (3R,10R,14S)-
diphenyl unsaturated ketal aminolactam (10). These compounds
were prepared from diphenyl diketal dilactam 1c (66.4 mg,
0.15 mmol), LiAlH4 (57 mg, 1.5 mmol, 10 equiv.), NEt3 (1 M solution
in THF, 30 mL, 30 mmol, 0.02 equiv./LAH), and THF (19 mL,
csubstrat ¼ 8.0 ꢀ 10ꢁ3 M) according to the general procedure (reac-
tion time 9 h). Excess hydride was destroyed by adding H2O
(270
60:40) afforded diketal aminolactam 4c (18.9 mg, 44
diketal diamine 7c (5.2 mg, 12.6 mol, 10%) and unsaturated ketal
aminolactam 10 (4.0 mg, 10.1 mol, 8%) with 84% conversion rate.
Data of 4c, 7c, and 10 were given previously [12a].
mL, 15 mmol). Chromatography (Et2O/MeOH, gradient 100:0–
mmol, 35%),
m
m
importance of phenyl groups susceptible to induce
p–p stacking
effects with aromatic units of the CXCR4 receptor. The development
and biological evaluation of new analogs incorporating phenyl
substituents would enable us to test this hypothesis.
6.1.2.3. (3S,7S,10S,14R)-Dimethyl
diketal
aminolactam
(5b),
(3S,7S,10S,14R)-dimethyl diketal diamine (8b), and (3S,10S,14R)-
dimethyl unsaturated ketal aminolactam (11). These compounds
were prepared from methyl diketal dilactam 2b (63.7 mg,
0.20 mmol), LiAlH4 (76 mg, 2.0 mmol, 10 equiv.), NEt3 (1 M solution
6. Experimental
6.1. Chemistry
in THF, 20 mL, 20 mmol, 0.01 equiv./LAH), and THF (25 mL,
csubstrat ¼ 8.0 ꢀ 10ꢁ3 M) according to the general procedure (reac-
6.1.1. General remarks
tion time 18 h). Excess hydride was destroyed by adding H2O
Solvents were obtained dried as follows: tetrahydrofuran (THF)
was distilled from benzophenone ketyl, CH2Cl2 and CH3CN were
refluxed over and distilled from CaH2, CH3OH was distilled from
magnesium. The organic layers were dried with Mg2SO4. Reactions
were monitored by thin layer chromatography (TLC) on aluminium
plates precoated with silica gel (Merck 60 F254) and spots visualized
by UV light or by spraying with a ceric sulphate and ammonium
molybdate acid solution. Flash chromatography was carried out
with silica gel (Merck 0.040–0.063 mm). Optical rotations were
measured at the sodium D line (589 nm) using a 1 dm quartz cell
with a JASCO DIP-370 apparatus. IR spectra were scanned with
a Perkin–Elmer 881 spectrophotometer (nmax in cmꢁ1). 1D (1H and
13C-J MOD) and 2D (COSY 1H-1H, HSQC 1H-13C) NMR spectra were
recorded with a Bruker Avance 400 spectrometer equipped with
(360
0:100) afforded diketal aminolactam 5b (15.0 mg, 49.3
diketal diamine 8b (6.1 mg, 21.1 mol, 12%) and unsaturated ketal
aminolactam 11 (0.9 mg, 3.5 mol, 2%) with 88% conversion rate.
Data of 5b, 8b, and 11 were given previously [12b].
m
L, 20 mmol). Chromatography (Et2O/MeOH, gradient 100:0–
m
mol, 28%),
m
m
6.1.2.4. (3S,7R,10S,14R)-Dimethyl
diketal
aminolactam
(5c),
(3S,7R,10S,14R)-dimethyl diketal diamine (8c) and (3S,10S,14R)-
dimethyl unsaturated ketal aminolactam (11). These compounds
were prepared from methyl diketal dilactam 2c (63.7 mg,
0.20 mmol), LiAlH4 (45.5 mg, 1.2 mmol, 6 equiv.), NEt3 (1 M solu-
tion in THF, 12 mL, 12 mmol, 0.01 equiv./LAH), and THF (50 mL,
csubstrat ¼ 4.0 ꢀ 10ꢁ3 M) according to the general procedure (reac-
tion time 9 h). Excess hydride was destroyed by adding H2O (216 mL,
a 1H/13C probe (1H: 400 MHz; 13C: 100 MHz), and 2D HMBC 1H-13
NMR spectra with an Avance 500 apparatus long dist 7.7 Hz (1H:
500 MHz; 13C: 125 MHz). Chemical shifts
were noted in parts per
C
12 mmol). Chromatography (Et2O/MeOH, gradient 100:0–0:100)
afforded diketal aminolactam 5c (4.4 mg, 14.4 mol, 12%), diketal
diamine 8c (7.7 mg, 26.4 mol, 22%) and unsaturated ketal ami-
nolactam 11 (0.3 mg,1.2 mol,1%) with 60% conversion rate. Data of
m
d
m
million (ppm) and splitting patterns designated as follows: singlet
(s), doublet (d), triplet (t), quartet (q), doubled doublet (dd),
doublet of doublet of doublet (ddd), doubled triplet (dt), multiplet
(m), broad signal (br. s). The solvent (CDCl3) was taken as internal
m
5c, 8c, and 11 were given previously [12b].
6.1.2.5. (7S*,14R*)-Diketal aminolactam (6b), (7S*,14R*)-diketal
diamine (9b), and 14R*-unsaturated ketal aminolactam (12). These
compounds were prepared from diketal dilactam 3b (72.6 mg,
0.25 mmol), LiAlH4 (95 mg, 2.5 mmol, 10 equiv.), NEt3 (1 M solu-
reference [
d
¼ 7.27 ppm for 1H NMR and 77.1 ppm (central line) for
13C NMR]. Proton and carbon atoms were assigned according to the
numbering indicated in Fig. 2 and Scheme 1. Mass spectra were
recorded with either an HP 5989B (CI) or a micro Q-TOF Waters
(ESI) apparatus.
tion in THF, 50 mL, 50 mmol, 0.02 equiv./LAH), and THF (50 mL,
csubstrat ¼ 5.0 ꢀ 10ꢁ3 M) according to the general procedure
(reaction time 9 h). Excess hydride was destroyed by adding H2O
6.1.2. Diketal aminolactams: general procedure
To a suspension of LiAlH4 (10 equiv.) in dry THF (0.5 mL for
1 mmol of LAH) was added NEt3 (0.01–0.04 equiv./LAH), and drop-
(450
0:100) afforded diketal aminolactam 6b (14.5 mg, 52.6
27%), diketal diamine 9b (8.2 mg, 31.2 mol, 16%) and unsaturated
mol, 2%) with 78% conversion
m
L, 25 mmol). Chromatography (Et2O/MeOH, gradient 100:0–
mmol,
m
wise
a
solution of diketal dilactam (1 equiv.) in dry THF
ketal aminolactam 12 (1.0 mg, 4.0 m
(csubstrate ¼ 3–14 ꢀ10ꢁ3 M). The reaction mixture was heated at
reflux while stirring for 9–18 h. Excess hydride was destroyed by
adding H2O (10 equiv./LAH). The precipitates were filtered and
washed with THF and CH2Cl2. The organic layer was dried with
MgSO4 and evaporated in vacuo. Purification by flash
rate.
Data for 6b: Rf ¼ 0.50 (MeOH/NH4OH: 99.5/0.5, 2 elutions). IR
(CHCl3): nmax 3450, 3420, 3340 (NH); 1660 (CO). 1H NMR and COSY
1H-1H (400 MHz, CDCl3): dH 1.89 (dddd, 1H, H-13B, 2J ¼ 14.5 Hz,
3J ¼ 7.6, 2.7, 2.4 Hz), 1.99 (dddd, 1H, H-13A, 2J ¼ 14.5 Hz, 3J ¼ 8.7, 8.1,

R. Affani et al. / European Journal of Medicinal Chemistry 44 (2009) 3138–3146
3143
2.8 Hz), 2.53 (dd, 1H, H-6B, 2J ¼ 14.6 Hz, 3J ¼ 1.9 Hz), 2.56 (br. s, 1H,
NH amine), 2.70 (dd, 1H, H-6A, 2J ¼ 14.6 Hz, 3J ¼ 8.6 Hz), 2.75 (ddd,
1H, H-12B, 2J ¼ 11.5 Hz, 3J ¼ 8.7, 2.7 Hz), 2.81 (ddd, 1H, H-10B,
2J ¼ 12.6 Hz, 3J ¼ 5.3, 2.6 Hz), 2.89 (ddd, 1H, H-12A, 2J ¼ 11.5 Hz,
3J ¼ 7.6, 2.8 Hz), 2.91 (ddd, 1H, H-10A, 2J ¼ 12.6 Hz, 3J ¼ 8.5, 2.8 Hz),
3.33 (s, 3H, OCH3), 3.36 (s, 3H, OCH3), 3.38 (dddd, 1H, H-3B,
2J ¼ 13.7, 3J ¼ 6.4, 5.2, 2.4 Hz), 3.51 (ddd, 1H, H-2B, 2J ¼ 9.9 Hz,
3J ¼ 7.9, 2.4 Hz), 3.55 (ddd, 1H, H-9B, 2J ¼ 9.9 Hz, 3J ¼ 8.5, 2.6 Hz),
3.63 (dddd, 1H, H-3A, 2J ¼ 13.7 Hz, 3J ¼ 7.9, 5.6, 2.6 Hz), 3.82 (ddd,
1H, H-2A, 2J ¼ 9.9 Hz, 3J ¼ 6.4, 2.6 Hz), 3.96 (ddd, 1H, H-9A,
2J ¼ 9.9 Hz, 3J ¼ 5.3, 2.8 Hz), 4.62 (dd, 1H, H-14, 2J ¼ 8.1, 2.4 Hz), 4.80
(dd, 1H, H-7, 3J ¼ 8.6, 1.9 Hz), 7.00 (br. s, 1H, NH lactam). 13C NMR
and HSQC (100 MHz, CDCl3): dC 31.6 (C-13), 39.3 (C-3), 41.1 (C-6),
45.7 (C-12), 49.3 (C-10), 53.1 (OCH3), 53.6 (OCH3), 64.1 (C-2), 65.3
(C-9), 101.2 (C-7), 104.0 (C-14), 169.0 (CO). MS (ESIþ): m/z (%) 299
(28) [M þ Na]þ, 277 (100) [M þ H]þ, 245 (87) [M þ H-CH3OH]þ.
HRMS (ESIþ): calcd. for C12H25N2O5 [M þ H]þ 277.1763, found
277.1767.
(Et2O/MeOH, gradient: 100:0–50:50) afforded bis-diketal amino-
lactam 13b (16.0 mg, 16.8 mol, 61%) and bromoxylyl diphenyl
m
diketal aminolactam 16b (5.4 mg, 8.8
mmol, 16%) with 92% conver-
sion rate.
25
Data for 13b: Rf ¼ 0.40 (Et2O/MeOH, 98:2, 3 elutions). [
a]
ꢁ68.0 (c 0.90, CHCl3). IR (CHCl3): nmax 3367 (NH), 1665 (CO). 1DH
NMR and COSY 1H-1H (400 MHz, CDCl3): dH 1.86 (m, 4H, 2H-13A,
2H-13B), 2.56 (dd, 2H, 2H-6B, 2J ¼ 16.1 Hz, 3J ¼ 1.5 Hz), 2.61 (dt, 2H,
2H-12B, 2J ¼ 12.8 Hz, 3J ¼ 4.9 Hz), 2.84 (dd, 2H, 2H-6A, 2J ¼ 16.1 Hz,
3J ¼ 8.7 Hz), 3.10 (s, 6H, 2 OCH3), 3.16 (s, 6H, 2 OCH3), 3.30 (d, 2H,
2H-15B, 2J ¼ 14.7 Hz), 3.34 (m, 2H, 2H-12A), 3.76 (dd, 2H, 2H-2B,
2J ¼ 9.8 Hz, 3J ¼ 3.2 Hz), 3.80 (d, 2H, 2H-15A, 2J ¼ 14.7 Hz), 3.92 (dd,
2H, 2H-9B, 2J ¼ 10.0 Hz, 3J ¼ 2.2 Hz), 3.97 (dd, 2H, 2H-10, 3J ¼ 7.3,
2.2 Hz), 4.10 (dd, 2H, 2H-2A, 2J ¼ 9.8 Hz, 3J ¼ 2.9 Hz), 4.11 (dd, 2H,
2H-9A, 2J ¼ 10.0 Hz, 3J ¼ 7.3 Hz), 4.79 (dd, 2H, 2H-7, 3J ¼ 8.7, 1.5 Hz),
4.84 (t, 2H, 2H-14, 3J ¼ 5.3 Hz), 5.28 (ddd, 2H, 2H-3, 3J ¼ 7.9, 3.2,
2.9 Hz), 7.28–7.42 (m, 24H, 24 Ar-H), 7.90 (d, 2H, 2 NH lactam,
3J ¼ 7.9 Hz). 13C NMR and HSQC (100 MHz, CDCl3): dC 31.1 (2C-13),
40.0 (2C-6), 44.7 (2C-12), 52.0 (2 OCH3), 52.3 (2C-3), 53.4 (2CH3),
53.5 (2C-15), 63.4 (2C-10), 66.4 (2C-9), 70.5 (2C-2), 100.1 (2C-7),
102.3 (2C-14), 126.7–129.4 (14 ArCH), 136.6 (2 ArC), 138.3 (2 ArC),
140.4 (2 ArC), 168.3 (2CO). MS (ESIþ): m/z (%) 981 (21) [M þ Na]þ,
959 (7) [M þ H]þ, 451 (100) [C24H32N2O5 þ Na]þ, 429 (84)
[C24H32N2O5 þ H]þ, 339 (42). HRMS (ESIþ): calcd. for C56H71N4O10
Data of 9b were given previously [12b].
Data for 12: Rf ¼ 0.60 (MeOH, 3 elutions). IR (CHCl3): nmax 3429
(NH), 1663 (CO), 1604 (C]C). 1H NMR and COSY 1H-1H (400 MHz,
CDCl3): dH 1.60 (br. s, 1H, NH amine), 1.82 (dddd, 1H, H-13B,
2J ¼ 14.3 Hz, 3J ¼ 7.2, 2.2, 2.1 Hz), 1.99 (dddd,1H, H-13A, 2J ¼ 14.3 Hz,
3J ¼ 9.3, 8.5, 2.3 Hz), 2.70 (ddd, 1H, H-12B, 2J ¼ 11.5, 3J ¼ 9.3, 2.1 Hz),
2.88 (m, 3H, H-10A, H-10B, H-12A), 3.28 (dddd, 1H, H-3B, 2J ¼ 11.0,
3J ¼ 9.0, 3.5, 2.5 Hz), 3.33 (s, 3H, OCH3), 3.47 (td, 1H, H-2B, 2J ¼ 10.0,
3J ¼ 2.5 Hz), 3.85 (ddt, 1H, H-3A, 2J ¼ 11.0, 3J ¼ 10.0, 3.5 Hz), 3.91
(dt, 1H, H-2A, 2J ¼ 10.0, 3J ¼ 3.5 Hz), 4.07 (ddd, 1H, H-9B, 2J ¼ 10.3,
3J ¼ 4.3, 4.1 Hz), 4.21 (ddd, 1H, H-9A, 2J ¼ 10.3, 3J ¼ 5.9, 4.9 Hz), 4.58
(dd, 1H, H-14, 3J ¼ 8.5, 2.2 Hz), 5.01 (d, 1H, H-6, 3J ¼ 6.9 Hz), 6.48 (d,
1H, H-7, 3J ¼ 6.9 Hz), 7.60 (br. s, 1H, NH lactam). MS (ESIþ): m/z (%)
267 (100) [M þ Na]þ, 245 (52) [M þ H]þ, 213 (98) [M þ H-CH3OH]þ.
HRMS (ESIþ): calcd. for C11H21N2O4 [M þ H]þ 245.1501, found
245.1514.
[M þ H]þ 959.5170, found 959.5210.
25
Data for 16b: Rf ¼ 0.60 (Et2O). [
a]
ꢁ56.6 (c 0.57, CHCl3). IR
D
(CHCl3) nmax 3360 (NH), 1660 (CO). 1H NMR and COSY 1H-1H
(400 MHz, CDCl3): dH 1.85 (m, 2H, H-13A, H-13B), 2.55 (ddd, 1H,
H-12B, 2J ¼ 11.8 Hz, 3J ¼ 4.7, 0.5 Hz), 2.57 (dd, 1H, H-6B, 2J ¼ 16.2 Hz,
3J ¼ 1.6 Hz), 2.83 (dd, 1H, H-6A, 2J ¼ 16.2 Hz, 3J ¼ 8.7 Hz), 3.11 (s, 3H,
7-OCH3), 3.14 (s, 3H,14-OCH3), 3.31 (d,1H, H-15B, 2J ¼ 15.0 Hz), 3.32
(ddd, 1H, H-12A, 2J ¼ 11.8 Hz, 3J ¼ 10.2, 6.5 Hz), 3.74 (dd, 1H, H-2B,
2J ¼ 10.0 Hz, 3J ¼ 3.3 Hz), 3.79 (d, 1H, H-15A, 2J ¼ 15.0 Hz), 3.89 (dd,
1H, H-9B, 2J ¼ 10.2 Hz, 3J ¼ 2.6 Hz), 3.96 (dd, 1H, H-10, 3J ¼ 8.4,
2.6 Hz), 4.09 (dd,1H, H-2A, 2J ¼ 10.0 Hz, 3J ¼ 2.7 Hz), 4.13 (dd,1H, H-
9A, 2J ¼ 10.2 Hz, 3J ¼ 8.4 Hz), 4.52 (s, 2H, CH2Br), 4.78 (dd, 1H, H-14,
3J ¼ 5.9, 5.3 Hz), 4.79 (dd, 1H, H-7, 3J ¼ 8.7, 1.6 Hz), 5.27 (ddd, 1H, H-
3, 3J ¼ 7.6, 3.3, 2.7 Hz), 7.22–7.42 (m, 14H, 14 Ar-H), 7.90 (d, 1H, NH
lactam, 3J ¼ 7.6 Hz). 13C NMR and HSQC (100 MHz, CDCl3): dC 31.1
(C-13), 33.6 (CH2Br), 40.1 (C-6), 44.4 (C-12), 52.1 (14-OCH3), 52.2
(C-3), 53.2 (7-OCH3), 53.4 (C-15), 63.6 (C-10), 66.3 (C-9), 70.6 (C-2),
100.0 (C-7), 102.3 (C-14), 127.0–129.4 (14 Ar-CH), 136.1 (Ar-C), 136.6
(Ar-C), 140.2 (Ar-C), 140.4 (Ar-C), 168.2 (CO). MS (ESIþ): m/z (%) 635
(9) [M þ Na (81Br)]þ, 633 (9) [M þ Na (79Br)]þ, 613 (98) [M þ H
6.1.2.6. (7R*,14R*)-Diketal aminolactam (6c) and (7R*,14R*)-diketal
diamine (9c). These compounds were prepared from methyl dike-
tal dilactam 3c (116.1 mg, 0.40 mmol), LiAlH4 (152 mg, 4.0 mmol,
10 equiv.), NEt3 (1 M solution in THF, 80 mL, 80 mmol, 0.02 equiv./
LAH), and THF (133 mL, csubstrate ¼ 3.0 ꢀ 10ꢁ3 M) according to the
general procedure (reaction time 10 h). Excess hydride was
destroyed by adding H2O (720
(Et2O/MeOH, gradient 100:0–0:100) afforded diketal aminolactam
6c (22.5 mg, 81.4 mol, 29%) and diketal diamine 9c (10.3 mg,
39.3 mol, 14%) with 70% conversion rate. Data of 6c and 9c were
mL, 40 mmol). Chromatography
m
m
(
81Br)]þ, 611 (100) [M þ H (79Br)]þ, 555 (16), 553 (16). HRMS (ESIþ):
given previously [12b].
calcd. for C32H40 79BrN2O5 [M þ H]þ 611.2121, found 611.2132.
6.1.3. Bis-diketal aminolactams: general procedure
6.1.3.2. Bis-(3R,7S,10R,14S)-diphenyl diketal aminolactam (13c) and
A suspension of diketal aminolactam,
a,
a
0-dibromo-p-xylene
(3R,7S,10R,14S)-bromoxylyl
(16c). These compounds were prepared from diphenyl diketal ami-
nolactam 4c (17.1 mg, 40.0 mol),
0-dibromo-p-xylene (6.3 mg,
24.0 mol, 0.6 equiv.), K2CO3 (6.6 mg, 48.0 mol, 1.2 equiv.), and
diphenyl
diketal
aminolactam
(0.5–0.6 equiv.) and K2CO3 (1.0–1.2 equiv.) in dry CH3CN
(csubstrate ¼ 50 ꢀ 10ꢁ3 M) was heated at reflux while stirring for
8–9 h. After evaporation of the solvent, the residue was dissolved in
CH2Cl2. The precipitate was filtered and washed with CH2Cl2. The
organic layer was dried with Mg2SO4 and evaporated in vacuo.
Purification by flash chromatography (silica gel, Et2O/MeOH) yiel-
ded bis-diketal aminolactams and small amounts of bromo diketal
aminolactams.
m
a,a
m
m
CH3CN (0.8 mL, csubstrate ¼ 50 ꢀ 10ꢁ3 M) according to the general
procedure (reaction time 8 h). Chromatography (Et2O/MeOH,
gradient100:0–50:50) affordedbis-diketalaminolactam13c (7.1 mg,
7.4
mmol, 37%) and bromoxylyl diphenyl diketal aminolactam 16c
(4.7 mg, 7.7
m
mol, 19%) with 100% conversion rate.
25
Data for 13c: Rf ¼ 0.33 (Et2O/MeOH, 98:2). [
a
]
ꢁ56.1 (c 0.25;
D
6.1.3.1. Bis-(3R,7R,10R,14S)-diphenyl diketal aminolactam (13b) and
CHCl3). IR (CHCl3): nmax 3436 and 3372 (NH), 1666 (CO). 1H NMR
and COSY 1H-1H (400 MHz, CDCl3): dH 1.87 (q, 4H, 4H-13,
3J ¼ 6.4 Hz), 2.59 (dd, 2H, 2H-6B, 2J ¼ 14.5 Hz, 3J ¼ 2.4 Hz), 2.63 (dd,
2H, 2H-6A, 2J ¼ 14.5 Hz, 3J ¼ 7.0 Hz), 2.76 (dt, 2H, 2H-12B,
2J ¼ 12.9 Hz, 3J ¼ 6.4 Hz), 3.11 (dt, 2H, 2H-12A, 2J ¼ 12.9 Hz,
3J ¼ 6.4 Hz), 3.19 (s, 12H, 4 OCH3), 3.43 (d, 2H, 2H-15B, 2J ¼ 14.3 Hz),
3.74 (d, 2H, 2H-15A, 2J ¼ 14.3 Hz), 3.83 (dd, 2H, 2H-9B, 2J ¼ 10.3 Hz,
(3R,7R,10R,14S)-bromoxylyl
(16b). These compounds were prepared from diphenyl diketal
aminolactam 4b (25.7 mg, 60.0 mol),
0-dibromo-p-xylene
mol,
diphenyl
diketal
aminolactam
m
a,a
(7.9 mg, 30.0 mmol, 0.5 equiv.), K2CO3 (8.3 mg, 60.0 m
1.0 equiv.), and CH3CN (1.2 mL, csubstrate ¼ 50 ꢀ 10ꢁ3 M) according
to the general procedure (reaction time 8 h). Chromatography

3144
R. Affani et al. / European Journal of Medicinal Chemistry 44 (2009) 3138–3146
3J ¼ 3.3 Hz), 3.84 (dd, 2H, 2H-2B, 2J ¼ 10.8 Hz, 3J ¼ 4.2 Hz,), 3.91 (dd,
2H, 2H-2A, 2J ¼ 10.8 Hz, 3J ¼ 4.9 Hz), 3.92 (dd, 2H, 2H-10, 3J ¼ 7.4,
3.3 Hz), 4.23 (dd, 2H, 2H-9A, 2J ¼ 10.3 Hz, 3J ¼ 7.4 Hz), 4.78 (dd, 2H,
2 H-7, 3J ¼ 7.0, 2.4 Hz), 4.79 (t, 2H, 2H-14, 3J ¼ 6.4 Hz), 5.24 (ddd, 2H,
2H-3, 3J ¼ 7.7, 4.9, 4.2 Hz), 7.13 (d, 2H, 2 NH lactam, 3J ¼ 7.7 Hz),
7.28–7.40 (m, 24H, 24 Ar-H). 13C NMR and HSQC (100 MHz, CDCl3)
dC 31.6 (2C-13), 41.3 (2C-6), 45.4 (2C-12), 52.4 (2CH3), 52.4 (2C-3),
54.1 (2 OCH3), 54.2 (2C-15), 62.8 (2C-10), 67.3 (2C-9), 69.7 (2C-2),
101.7 (2C-7), 102.2 (2C-14), 127.0–128.9 (24 Ar-CH), 138.3 (2 Ar-C),
138.5 (2 Ar-C), 139.6 (2 Ar-C), 168.7 (2CO). MS (ESIþ): m/z (%) 981
(23) [M þ Na]þ, 959 (100) [M þ H]þ, 613 (14), 499 (23), 491 (10).
HRMS (ESIþ): calcd. for C56H71N4O10 [M þ H]þ 959.5170, found
(2C-9), 71.4 (2C-2), 99.7 (2C-7), 102.3 (2C-14), 128.4 (4 Ar-CH), 138.8
(2 Ar-C), 167.9 (2CO). MS (APCIþ) m/z (%) 711 (100) [MHþ], 679 (30)
[M þ H-CH3OH]þ, 647 (7). HRMS (ESIþ): calcd. for C36H63N4O10
[M þ H]þ 711.4544, found 711.4576.
Data for 17b: Rf ¼ 0.55 (Et2O/MeOH, 50:50), 0.75 (MeOH/NH4OH,
25
99:1). [
a
]
þ19.0 (c 0.65, CHCl3). IR (CHCl3): nmax 3365 and 3290
D
(NH), 1662 (CO). 1H NMR and COSY 1H-1H (400 MHz, CDCl3): dH 1.02
(d, 3H, CH3, 3J ¼ 6.8 Hz), 1.25 (d, 3H, CH3, 3J ¼ 6.8 Hz), 1.84 (m, 2H, H-
13A, H-13B), 2.50 (dd,1H, H-6B, 2J ¼ 16.6 Hz, 3J ¼ 1.9 Hz), 2.54 (dt, 1H,
H-12B, 2J ¼ 12.5 Hz, 3J ¼ 4.1 Hz), 2.73 (dd, 1H, H-6A, 2J ¼ 16.6 Hz,
3J ¼ 9.0 Hz), 2.92 (dqd,1H, H-10, 3J ¼ 6.8, 6.0, 2.0 Hz), 3.15 (ddd,1H, H-
12A, 2J ¼ 12.5 Hz, 3J ¼ 11.0, 6.1 Hz), 3.25 (s, 3H, OCH3), 3.31 (s, 3H,
OCH3), 3.41 (d, 1H, H-15B, 2J ¼ 14.7 Hz), 3.44 (dd, 1H, H-2B,
2J ¼ 10.0 Hz, 3J ¼ 2.8 Hz), 3.55 (ABX, AB part, 2H, H-9A, H-9B, 2J ¼ 10.0,
3J ¼ 6.0, 2.0 Hz, Dn ¼ 11.1 Hz), 3.75 (dd, 1H, H-2A, 2J ¼ 10.0 Hz,
3J ¼ 1.8 Hz), 3.81(d,1H, H-15A, 2J ¼ 14.7 Hz), 4.26(m,1H, H-3), 4.50(s,
2H, CH2Br), 4.71 (dd, 1H, H-7, 3J ¼ 9.0, 1.9 Hz), 4.75 (dd, 1H, H-14,
3J ¼ 7.8, 3.2 Hz), 7.26 (d, 4H, 4 Ar-H, 3J ¼ 7.1 Hz), 7.46 (d,1H, NHlactam,
3J ¼ 8.1 Hz). 13C NMR and HSQC (100 MHz, CDCl3) dC 8.1 (CH3), 18.1
(CH3),30.9(C-13), 33.6(CH2Br),39.5(C-6), 44.3(C-12), 44.5(C-3),52.0
(OCH3), 53.0 (C-10), 54.0 (C-15), 54.3 (OCH3), 67.7 (C-2), 71.4 (C-9),
99.7 (C-7),102.3 (C-14),128.5–129.2 (4 Ar-CH),137.1 (Ar-C),145.0 (Ar-
C),168.0 (CO). MS (APCIþ): m/z (%) 489 (92) [M þ H (81Br)]þ, 487 (79)
[M þ H (79Br)]þ, 457 (57) [M þ H-CH3OH (81Br)]þ, 455 (36) [M þ H-
CH3OH (79Br)]þ, 431 (100), 429 (91), 385 (15), 127 (49). HRMS (ESIþ):
calcd for C22H36 79BrN2O5 [M þ H]þ 487.1807, found 487.1826.
959.5214.
20
Data for 16c: Rf ¼ 0.66 (Et2O/MeOH, 98:2). [
a
]
ꢁ77.2 (c 0.23,
D
CHCl3). IR (CHCl3): nmax 3435 and 3360 (NH), 1666 (CO). 1H NMR
and COSY 1H-1H (400 MHz, CDCl3): dH 1.85 (m, 2H, H-13A, H-13B),
2.60 (dd, 1H, H-6B, 2J ¼ 14.6 Hz, 3J ¼ 2.6 Hz), 2.66 (dd, 1H, H-6A,
2J ¼ 14.6 Hz, 3J ¼ 6.8 Hz), 2.74 (ddd, 1H, H-12B, 2J ¼ 13.3 Hz, 3J ¼ 6.8,
5.7 Hz), 3.15 (ddd, 1H, H-12A, 2J ¼ 13.3 Hz, 3J ¼ 7.5, 5.6 Hz), 3.18 (s,
3H, OCH3), 3.24 (s, 3H, OCH3), 3.45 (d, 1H, H-15B, 2J ¼ 14.7 Hz), 3.74
(d, 1H, H-15A, 2J ¼ 14.7 Hz), 3.79 (dd, 1H, H-9B, 2J ¼ 10.5 Hz,
3J ¼ 2.9 Hz), 3.83 (dd, H, H-2B, 2J ¼ 10.7 Hz, 3J ¼ 3.6 Hz), 3.91 (dd,1H,
H-10, 3J ¼ 7.7, 2.9 Hz), 3.92 (dd, 1H, H-2A, 2J ¼ 10.7 Hz, 3J ¼ 4.9 Hz),
4.25 (dd,1H, H-9A, 2J ¼ 10.5 Hz, 3J ¼ 7.7 Hz), 4.51 (s, 2H, CH2Br), 4.77
(dd, 1H, H-14, 3J ¼ 6.6, 4.0 Hz), 4.78 (dd, 1H, H-7, 3J ¼ 6.8, 2.6 Hz),
5.24 (ddd, 1H, H-3, 3J ¼ 7.7, 4.9, 3.6 Hz), 7.15 (d, 1H, NH lactam,
3J ¼ 7.7 Hz), 7.28–7.40 (m, 14H, 14 Ar-H). 13C NMR and HSQC
(100 MHz, CDCl3), HMBC (125 MHz, CDCl3):
d
31.7 (C-13), 33.7
6.1.3.4. Bis-(7S*,14R*)-diketal aminolactam (15b). This compound
(CH2Br), 41.2 (C-6), 45.3 (C-12), 52.5 (C-3), 52.6 (OCH3), 54.2 (C-15),
53.3 (OCH3), 63.1 (C-10), 67.3 (C-9), 69.8 (C-2), 101.8 (C-7), 102.2
(C-14), 127.0–129.2 (14 Ar-CH), 136.4 (Ar-C), 138.0 (Ar-C), 139.6 (Ar-
C), 140.6 (Ar-C), 168.6 (CO). MS (ESIþ): m/z (%) ¼ 635 (29) [M þ Na
was prepared from diketal aminolactam 6b (13.8 mg, 50.0
mmol),
a,
a
0-dibromo-p-xylene (6.6 mg, 25.0
mmol, 0.5 equiv.), K2CO3
(6.9 mg,
50.0
m
mol,
1.0 equiv.),
and
CH3CN
(1.0 mL,
csubstrate ¼ 50 ꢀ 10ꢁ3 M) according to the general procedure (reac-
(
81Br)]þ, 633 (28) [M þ Na (79Br)]þ, 613 (100) [M þ H (81Br)]þ, 611
tion time 9 h). Chromatography (Et2O/MeOH, gradient 100:0–
(98) [M þ H (79Br)]þ. HRMS (ESIþ): calcd. for C32H40 79BrN2O5
0:100) afforded bis-diketal aminolactam 15b (6.6 mg, 10.1 mmol,
[M þ H]þ 611.2121, found 611.2131.
48%) with 84% conversion rate. Rf ¼ 0.38 (MeOH/NH4OH, 99:1). IR
(CHCl3): nmax 3450 and 3369 (NH), 1656 (CO). 1H NMR and COSY
1H-1H (400 MHz, CDCl3): dH 1.91 (m, 4H, 4H-13), 2.48 (m, 2H, 2H-
10B), 2.52 (dd, 2H, 2H-6B, 2J ¼ 15.1 Hz, 3J ¼ 1.0 Hz), 2.58 (m, 2H,
2H-12B), 2.66 (dd, 2H, 2H-6A, 2J ¼ 15.1 Hz, 3J ¼ 7.8 Hz), 2.68 (m, 2H,
2H-10A), 2.88 (m, 2H, 2H-12A), 3.30 (s, 6H, 2 OCH3), 3.31 (s, 6H,
2 OCH3), 3.42 (d, 2H, 2H-15B, 2J ¼ 15.0 Hz), 3.50 (m, 4H, 4H-3), 3.51
(m, 2H, 2H-2B), 3.57 (ddd, 2H, 2H-9B, 2J ¼ 9.0 Hz, 3J ¼ 8.4, 1.0 Hz),
3.70 (d, 2H, 2H-15A, 2J ¼ 15.0 Hz), 3.73 (m, 2H, 2H-9A), 3.78 (m, 2H,
2H-2A), 4.70 (dd, 2H, 2H-7, 3J ¼ 7.8, 1.0 Hz), 4.73 (dd, 2H, 2H-14,
3J ¼ 10.4, 5.2 Hz), 7.25 (br. s, 2H, 2 NH lactam), 7.26 (d, 4H, 4 Ar-H,
3J ¼ 7.8 Hz). 13C NMR and HSQC (100 MHz, CDCl3,): dC 30.2 (2C-13),
38.8 (2C-3), 40.5 (2C-6), 49.9 (2C-12), 52.5 (2 OCH3), 53.4 (2 OCH3),
54.0 (2C-10), 57.9 (2C-15), 64.7 (2C-2), 65.2 (2C-9), 100.4 (2C-7),
101.4 (2C-14), 129.0 (4 Ar-CH), 140.3 (2 Ar-C), 169.0 (2CO). MS
(ESIþ): m/z (%) 693 (5) [M þ K]þ, 677 (76) [M þ Na]þ, 655 (100)
[M þ H]þ, 623 (4) [M þ H-CH3OH]þ. HRMS (ESIþ): calcd. for
C32H55N4O10 [M þ H]þ 655.3918, found 655.3931.
6.1.3.3. Bis-(3S,7S,10S,14R)-dimethyl diketal aminolactam (14b) and
(3S,7S,10S,14R)-bromoxylyl
(17b). These compounds were prepared from dimethyl diketal
aminolactam 5b (15.2 mg, 50.0 mol),
0-dibromo-p-xylene
(7.9 mg, 30.0 mol, 0.6 equiv.), K2CO3 (8.3 mg, 60.0 mol,
dimethyl
diketal
aminolactam
m
a,a
m
m
1.2 equiv.), and CH3CN (1.0 mL, csubstrate ¼ 50 ꢀ 10ꢁ3 M) according to
the general procedure (reaction time 9 h). Chromatography (Et2O/
MeOH, gradient 100:0–50:50) afforded bis-dimethyl diketal ami-
nolactam 14b (11.5 mg, 16.2 mmol, 72%) and bromoxylyl dimethyl
diketal aminolactam 17b (1.3 mg, 2.7 mmol, 6%) with 90% conver-
sion rate.
Data for 14b: Rf ¼ 0.35 (Et2O/MeOH, 50:50), 0.45 (MeOH/
25
NH4OH, 99:1). [
a
]
þ 55.2 (c 0.69, CHCl3). IR (CHCl3):
n 3365 (NH),
D
1653 (CO). 1H NMR, COSY 1H-1H (400 MHz, CDCl3): dH 1.02 (d, 6H,
2CH3, 3J ¼ 6.9 Hz), 1.24 (d, 6H, 2CH3, 3J ¼ 6.7 Hz), 1.86 (m, 4H,
4H-13), 2.49 (dd, 2H, 2H-6B, 2J ¼ 16.8 Hz, 3J ¼ 1.9 Hz), 2.53 (dt, 2H,
2H-12B, 2J ¼ 12.5 Hz, 3J ¼ 3.9 Hz), 2.72 (dd, 2H, 2H-6A, 2J ¼ 16.8 Hz,
3J ¼ 9.0 Hz), 2.90 (qdd, 2H, 2H-10, 3J ¼ 6.9, 6.3, 2.2 Hz), 3.16 (ddd,
2H, 2H-12A, 2J ¼ 12.5 Hz, 3J ¼ 11.5, 5.1 Hz), 3.27 (s, 6H, 2 OCH3), 3.32
(s, 6H, 2 OCH3), 3.35 (d, 2H, 2H-15B, 2J ¼ 14.3 Hz), 3.45 (dd, 2H, 2H-
2B, 2J ¼ 9.7 Hz, 3J ¼ 3.1 Hz), 3.54 (ABX system, AB part, 4H, 2H-9A,
2H-9B, 2J ¼ 10.0 Hz, 3J ¼ 6.3, 2.2 Hz, Dn ¼ 13.4 Hz), 3.76 (dd, 2H, 2H-
2A, 2J ¼ 9.7 Hz, 3J ¼ 2.0 Hz), 3.81 (d, 2H, 2H-15A, 2J ¼ 14.3 Hz), 4.27
(m, 2H, 2H-3), 4.73 (dd, 2H, 2H-7, 3J ¼ 9.0, 1.9 Hz), 4.81 (dd, 2H,
2H-14, 3J ¼ 8.7, 2.6 Hz), 7.26 (d, 4H, 4 ArH, 3J ¼ 7.1 Hz), 7.48 (d, 1H,
NH lactam, 3J ¼ 8.6 Hz). 13C NMR and HSQC (100 MHz, CDCl3): dC 8.0
(2CH3), 18.1 (2CH3), 30.8 (2C-13), 39.4 (2C-6), 44.4 (2C-12), 44.6
(2C-3), 51.9 (2 OCH3), 53.8 (2C-10), 53.9 (2C-15), 54.4 (2 OCH3), 67.8
6.1.3.5. Bis-(7R*,14R*)-diketal aminolactam (15c). This compound
was prepared from diketal aminolactam 6c (16.6 mg, 60.0
mmol),
a,
a
0-dibromo-p-xylene (9.5 mg, 36.0
mmol, 0.6 equiv.), K2CO3
(10.0 mg,
72.0
m
mol,
1.2 equiv.),
and
CH3CN
(1.2 mL,
csubstrate ¼ 50 ꢀ 10ꢁ3 M) according to the general procedure (reac-
tion time 9 h). Chromatography (Et2O/MeOH, gradient 100:0–
0:100) afforded bis-diketal aminolactam 15c (9.4 mg, 14.3 mmol,
55%) with 87% conversion rate. Rf ¼ 0.35 (MeOH). IR (CHCl3): nmax
3450 and 3350 (NH),1660 (CO). 1H NMR and COSY 1H-1H (400 MHz,
CDCl3): dH 1.88 (m, 4H, 4H-13), 2.53 (m, 6H, 2H-6B, 2H-10B,
2H-12B), 2.64 (m, 4H, 2H-6A, 2H-10A), 2.90 (dt, 2H, 2H-12A,
2J ¼ 12.0 Hz, 3J ¼ 6.0 Hz), 3.21 (dddd, 2H, 2H-3B, 2J ¼ 11.2 Hz,

R. Affani et al. / European Journal of Medicinal Chemistry 44 (2009) 3138–3146
3145
3J ¼ 7.7, 3.0, 1.0 Hz), 3.30 (s, 12H, 4 OCH3), 3.40 (d, 2H, 2H-15B,
2J ¼ 14.0 Hz), 3.50 (ddd, 2H, 2H-9B, 2J ¼ 8.1 Hz, 3J ¼ 7.3 Hz, 1.0 Hz),
3.57 (ddd, 2H, 2H-2B, 2J ¼ 9.0 Hz, 3J ¼ 8.4, 1.0 Hz), 3.72 (d, 2H, 2H-
15A, 2J ¼ 14.0 Hz), 3.76 (m, 4H, 2H-2A, 2H-9A), 3.81 (m, 2H, 2H-3A),
4.66 (dd, 2H, 2H-7, 3J ¼ 6.8, 1.0 Hz), 4.72 (t, 2H, 2H-14, 3J ¼ 5.2 Hz),
7.16 (br. s, 2H, 2 NH lactam), 7.26 (d, 4H, 4 ArH, 3J ¼ 8.0 Hz). 13C NMR
and HSQC (100 MHz, CDCl3): dC 30.9 (2C-13), 39.2 (2C-3), 40.8 (2C-
6), 50.2 (2C-12), 52.6 (2 OCH3), 53.6 (2 OCH3), 54.1 (2C-10), 58.2
(2C-15), 66.0 (2C-9, 2C-2), 100.9 (2C-7), 102.2 (2C-14), 129.1 (4 Ar-
CH), 140.2 (2C-16), 169.1 (2CO). MS (ESIþ): m/z (%) 693 (19)
[M þ K]þ, 677 (8) [M þ Na]þ, 655 (100) [M þ H]þ, 627 (4) [M þ H-
CH3OH]þ. HRMS (ESIþ): calcd. for C32H55N4O10 [M þ H]þ 655.3918,
found 655.3887.
Data for 19c: Rf ¼ 0.25 (MeOH/NH4OH, 94:4, 2 elutions). IR
(CHCl3): nmax 3300 (NH). NMR 1H (400 MHz, CDCl3): dH 1.70–2.10
(m,14H, 2H-6B, 2H-13B, 2H-6A, 2H-13A, 2H-5B, 2H-12B, 2NH), 2.54
(m, 4H, 2H-3B, 2H-10B), 2.70 (m, 4H, 2H-3A, 2H-10A), 3.21 (d, 2H,
2H-15B, 2J ¼ 14.5 Hz), 3.23 (m, 4H, 2H-5A, 2H-12A), 3.30 (s, 6H, 2
OCH3), 3.38 (s, 6H, 2 OCH3), 3.60–3.71 (m, 4H, 2H-2B, 2H-9B), 3.86–
3.93 (m, 4H, 2H-2A, 2H-9A), 3.98 (d, 2H, 2H-15A, 2J ¼ 14.5 Hz), 4.51
(ddd, 2H, 2H-7, 3J ¼ 8.0, 4.7 Hz, 4J ¼ 2.4 Hz), 4.58 (ddd, 2H, 2H-14,
3J ¼ 7.4, 2.3 Hz, 4J ¼ 1.7 Hz), 7.30 (d, 4H, 4 Ar-H, 3J ¼ 6.6 Hz). 13C NMR
(100 MHz, CDCl3): dC 30.2 (2C-13), 31.7 (2C-6), 43.6 (2C-5), 47.4 (2C-
3), 48.3 (2C-12), 52.5 (2C-10), 52.8 (2 OCH3), 54.8 (2 OCH3), 57.9
(2C-15), 63.3 (2C-2), 67.1 (2C-9), 102.4 (2C-14), 103.7 (2C-7), 129.7
(4 Ar-CH),140.3 (2 Ar-C). MS (ESIþ): m/z (%) 649 (40) [M þ Na]þ, 627
(100) [M þ H]þ. HRMS (ESIþ): calcd. for [M þ H]þ 627.4333, found
627.4329.
6.1.4. Bis-diketal diamines: general procedure
To a solution of bis-diketal aminolactam (1 equiv.) in dry THF
(csubstrate ¼ 60 ꢁ 100 ꢀ 10ꢁ3 M), was added, dropwise while stirring,
a solution of LiAlH4 (1 M solution in THF, 10 equiv.) and NEt3
(0.02 equiv./LAH). The reaction mixture was heated at reflux for 6–
9 h. Excess hydride was destroyed by adding H2O (10 equiv./LAH).
The precipitates were filtered and washed with THF and CH2Cl2.
The organic layer was dried with MgSO4 and evaporated in vacuo.
Purification by flash chromatography (silicagel, Et2O/MeOH and
MeOH/NH4OH) yielded bis-diketal diamines.
6.2. Biological assays
6.2.1. Anti-HIV activity
The cell cultures of MT-4 and CEM-SS cells were maintained at
37 ꢂC in a 5% CO2 atmosphere in RPMI 1640 medium supplemented
with 10% foetal calf serum (FCS) containing penicillin, streptomycin
and glutamine. MT-4 cells were infected with HIV-1IIIB isolate at 5
TCID50 (Tissue Culture Infective Dose) and CEM-SS cells with HIV-1
LAI isolate at 20 TCID50. After 30 min of adsorption, the infected
cells were washed to remove unadsorbed virus particles and
resuspended in RPMI at the final concentration of 105 cells/mL, in
the presence of various dilutions of the test compounds. After 5
days, virus production was measured as already described [16]. The
6.1.4.1. Bis-(3S,7S,10S,14R)-dimethyl diketal diamine (18b). This
compound was prepared from bis-dimethyl diketal aminolactam
14b (11.4 mg, 16 mmol), LiAlH4 (1 M THF solution, 160 mL, 160 mmol,
10 equiv.), NEt3 (1 M THF solution, 3.2 mL, 3.2 mmol, 0.02 equiv./
LAH), THF (160
general procedure (reaction time 9 h). Excess hydride was destroyed
m
L, csubstrate ¼ 100 ꢀ 10ꢁ3 M) according to the
50% inhibitory concentration (IC50) was derived from the
computer-generated median effect plot of the dose-effect data [17].
The cytotoxicity of the drugs was evaluated in parallel by incu-
bating uninfected cells in the presence of ranging concentrations of
anti-viral products. Cell viability was measured by means of
a colorimetric reaction based on the capacity of mitochondrial
dehydrogenase of living cells to reduce 3-(4,5-dimethylthiazol-2-
yl)-2,5-diphenyltetrazolium bromide (MTT) into formazan, the
quantity of which was determined by the optical density at 540 nm
[18]. The 50% cytotoxic concentration (CC50) was calculated with
the program used for the determination of IC50. The CEM-SS cells
were obtained from P. Nara through the AIDS Research and Refer-
ence Reagent Program, Division of AIDS, NIAID, NIH.
by adding H2O (29
gradient 100:0–0:100 and MeOH/NH4OH, gradient 100:0–90:10)
afforded bis-dimethyl diketal diamine 18b (2.9 mg, 4.2 mol, 26%)
and dimethyl diketal diamine 8b (0.8 mg, 2.8 mol, 9%).
mL, 1.6 mmol). Chromatography (Et2O/MeOH,
m
m
Data for 18b: Rf ¼ 0.20 (MeOH/NH4OH, 97:3). IR (CHCl3): nmax
3340 (NH). 1H NMR and COSY 1H-1H (400 MHz, CDCl3): dH 1.01 (d,
6H, 2CH3, 3J ¼ 6.8 Hz), 1.10 (d, 6H, 2CH3, 3J ¼ 6.6 Hz), 1.81 (m, 4H,
4H-13), 1.92 (m, 4H, 4H-6), 2.05 (br. s, 2H, 2 NH amine), 2.53 (dt, 2H,
2H-12B, 2J ¼ 12.5 Hz, 3J ¼ 5.1 Hz), 2.70 (m, 2H, 2H-5B), 2.90 (m, 6H,
2H-3, 2H-5A, 2H-10), 3.07 (ddd, 2H, 2H-12A, 2J ¼ 12.6 Hz, 3J ¼ 9.5,
6.2 Hz), 3.28 (s, 6H, 2 OCH3), 3.32 (s, 6H, 2 OCH3), 3.45 (d, 2H, 2H-
15B, 2J ¼ 14.4 Hz), 3.46 (dd, 2H, 2H-2B, 2J ¼ 10.0 Hz, 3J ¼ 2.4 Hz),
3.48 (td, 2H, 2H-9B, 2J ¼ 10.0 Hz, 3J ¼ 6.0 Hz), 3.61 (td, 2H, 2H-9A,
2J ¼ 10.0 Hz, 3J ¼ 2.5 Hz), 3.66 (dd, 2H, 2H-2A, 2J ¼ 10.0 Hz,
3J ¼ 2.3 Hz), 3.75 (d, 2H, 2H-15A, 2J ¼ 14.4 Hz), 4.58 (dd, 2H, 2H-7,
3J ¼ 6.4, 3.8 Hz), 4.77 (dd, 2H, 2H-14, 3J ¼ 7.1, 1.8 Hz), 7.28 (d, 4H, 4
ArH, 3J ¼ 6.8 Hz). 13C NMR and HSQC (100 MHz, CDCl3), HMBC
(125 MHz, CDCl3): dC 9.7 (2CH3), 16.0 (2CH3), 32.0 (2C-6, 2C-13),
42.2 (2C-5), 44.4 (2C-12), 52.8 (2C-3), 52.9 (2 OCH3), 54.3 (2 OCH3),
54.5 (2C-10), 54.6 (2C-15), 68.3 (2C-2), 70.0 (2C-9), 101.8 (2C-14),
103.4 (2C-7), 128.3 (4 Ar-CH), 139.2 (2 Ar-C). MS (APCI) m/z (%): 705
(9) [M þ Na] þ, 683 (100) [M þ H]þ, 655 (7), 613 (8). HRMS (ESIþ):
calcd. for C36H67N4O8 [M þ H]þ 683.4959, found 683.4955.
6.2.2. Anti-tumoral activity
The cell cultures were maintained at 37 ꢂC under 5% CO2
atmosphere for 72 h, in either RPMI 1640 medium (cell lines:
SF268, HL60, MCF7, MCF7R, HCT116, A549, PC3, OV3) or DMEM-
Glutamax medium (cell lines: KB, HepG2), supplemented in both
cases with 10% foetal calf serum (FCS) containing penicillin, strep-
tomycin, fungizon and glutamine. Cells were plated at a density of
500–800 cells in 200 mL culture medium in each well of 96-well
microplates and allowed to adhere for 24 h before treatment with
test drugs in DMSO solution (final concentration: 10ꢁ5 M). Cell
viability was assayed after 3 h drug exposure with tetrazolium
compound
[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethox-
6.1.4.2. Bis-(7R*,14R*)-diketal diamine (19c). This compound was
yphenyl)-2-(4-sulphophenyl)-2H-tetrazolium salt, MTS, Promega
Corp.] using the Cell Titer 96 Aqueous One Solution Cell Prolifera-
tion Assay kit (Promega Corp) according to the manufacturer’s
instructions. Absorbance was measured at 490 nm with a micro-
plate reader.
prepared from bis-diketal aminolactam 15c (10.5 mg, 16
mmol),
LiAlH4 (1 M THF solution, 160 L, 160 mol, 10 equiv.), NEt3 (1 M
m
m
THF solution, 3.2 mL, 3.2 mmol, 0.02 equiv./LAH), THF (267 mL,
csubstrate ¼ 60 ꢀ 10ꢁ3 M) according to the general procedure (reac-
tion time 6 h). Excess hydride was destroyed by adding H2O (29 mL,
1.6 mmol). Chromatography (Et2O/MeOH, gradient 100:0–0:100
and MeOH/NH4OH, gradient 100:0–70:30) afforded bis-diketal
Acknowledgements
diamine 19c (2.2 mg, 3.5
(0.8 mg, 3.0 mol, 9%).
mmol, 22%) and diketal diamine 9c
The authors thank R. Egrot for the 2D HMBC NMR spectra, B.
´
Legeret for the mass spectra (MS and HRMS), G. Laumond for the
m

3146
R. Affani et al. / European Journal of Medicinal Chemistry 44 (2009) 3138–3146
A. Mantovani, P. Allavena, Cancer Res. 64 (2004) 8420–8427;
(c) M.C.P. Smith, K.E. Luker, J.R. Garbow, J.L. Prior, E. Jackson, D. Piwnica-
Worms, G.D. Luker, Cancer Res. 64 (2004) 8604–8612;
(d) H. Kajiyama, K. Shibata, M. Terauchi, K. Ino, A. Nawa, F. Kikkawa, Int. J.
Cancer 122 (2008) 91–99.
anti-HIV assays, and the Centre National de la Recherche Scienti-
fique (CNRS), the Agence Nationale de Recherche sur le Sida (ANRS)
and the Plan Auvergne Cancer (PAC) for financial supports.
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