Antiviral activity and molecular geometry of some new symmetrical tris(aminoalkyl)amine derivatives

Article abstract of DOI:10.1248/cpb.c16-00682

We report the preparation of new tripodal receptor-type C₃- and C_S-symmetrical molecules constructed on a tris(2-aminoethyl)amine (TAEA) template. Both the anti-herpes simplex virus type 1 (anti-HSV-1) activity and cytotoxic activity

Full text of DOI:10.1248/cpb.c16-00682

Vol. 64, No. 12
Chem. Pharm. Bull. 64, 1769–1780 (2016)
1769
Note
Antiviral Activity and Molecular Geometry of Some New Symmetrical
Tris(aminoalkyl)amine Derivatives
Nobuko Mibu,a Kazumi Yokomizo,b Kana Murakami,a Yurika Ono,a Masato Ishimaru,a
,a
Marie Otsubo,a Hiroshi Inao,b Yutaro Ono,b Jian-Rong Zhou,b and Kunihiro Sumoto*
a Faculty of Pharmaceutical Sciences, Fukuoka University; 8–19–1 Nanakuma, Jonan-ku, Fukuoka 814–0180,
b
Japan: and Faculty of Pharmaceutical Sciences, Sojo University; 4–22–1 Ikeda, Kumamoto 860–0082, Japan.
Received August 26, 2016; accepted September 28, 2016
We report the preparation of new tripodal receptor-type C3- and CS-symmetrical molecules constructed
on a tris(2-aminoethyl)amine (TAEA) template. Both the anti-herpes simplex virus type 1 (anti-HSV-1)
activity and cytotoxic activity of synthesized receptor-type derivatives were evaluated in order to find a
characteristic structural feature for these bioactivities of compounds. Among the compounds of synthesized
symmetrical TAEA-related derivatives, compound 13k showed high anti-HSV-1 activity (50% effective con-
centration (EC50)=16.7 µM) and low cytotoxicity (50% cytotoxic concentration (CC50)=>200µM). The presence
of a hydrogen bond donor proton in the molecule is thought to be an important structural factor for express-
ing potential anti-HSV-1 activities.
Key words tris(2-aminoethyl)amine; C3 symmetry; CS symmetry; anti-herpes simplex virus type 1; tripodal
receptor-type; plaque reduction assay
It is well known that many types of receptors or functional-
ized proteins on membranes in a native state often have a high
Results
Synthesis of New Target Derivatives of Symmetrical
order of symmetrical interfaces.1) C3- or C2-symmetrical mole- Tris(aminoalkyl)amines (TAEA and TAPA) Our strategies
cules have frequently been found in various synthetic biologi- for the synthesis of target TAEA and TAPA derivatives by
cally active compounds.2) Aiming to develop new synthetic using TAEA (1) or TAPA (17) as a starting material are sum-
bioactive compounds, we targeted such symmetric geometrical marized in Chart 1.
molecules constructed on a symmetrical template.3–5)
The pathway for the synthesis of C3-symmetrical tripo-
In our previous studies on bioactive compounds in the tri- dal receptor-type TAEA derivatives (3, 5, 7, 8) is shown in
podal receptor-type tris(2-aminoethyl)amine (TAEA) series, Chart 1. All reductive amination reactions with various aryl
we found that some of the new C3- and CS-symmetrical de- aldehydes (2) were performed as one-pot reactions, and the
rivatives synthesized previously showed significant bioactivi- desired amine-type TAEA derivatives (3) were obtained in a
ties against herpes simplex virus type 1 (HSV-1) or cytotoxic manner similar to that for the preparation of compound 3a.13)
activities to Vero cells.6) Regarding amide-type TAEA deriva- Acetylation of compounds 3 with acid anhydride (4) gave new
tives, we have reported that some symmetrical molecules have amide-type TAEA derivatives (5). Urea-type TAEA deriva-
selective carbohydrate recognition properties.7) These mid-size tives (7, 8) were also obtained by addition reactions of amine-
molecules that show lectin-like carbohydrate recognition prop- type derivatives (3) or TAEA (1) to iso(thio)cyanate (6). All
erties might have interesting functions as new ligands or drug of these target C3-symmetrical TAEA derivatives (3, 5, 7, 8)
candidates. Considering the formation of strong drug-host in- were obtained in moderate to excellent yields (44–95%) (see
teraction for host sugar recognition by multivalent molecules,
attempts to synthesize such non-peptide mid-size molecules
are also thought to be valid in the search for bioactive new
seeds.5,8)
Since it is well known that many amide functionalities usu-
ally have the ability to form a strong hydrogen bond, and it
has also often been reported that some molecules derived from
TAEA behave as acceptors for an ion or a small molecule,11,12)
we introduced a few amide functionalities into a tripodal
receptor-type TAEA template. These molecular modifications
may lead to new bioactive symmetrical molecules that have
good properties for non-covalent interaction and bioactivities.
Here, we describe the synthesis of some new non-peptide
molecules with amine or amide-relating functionalities con-
structed on a symmetrical tris(2-aminoethyl)amine (TAEA)
template and on a spread tris(3-aminopropyl)amine (TAPA)
template. The results of biological evaluations of the synthe-
sized derivatives are also described (Fig. 1).
Fig. 1. Symmetrical Target Molecules Constructed on Triaminoalkyl-
Substituted Amines
*To whom correspondence should be addressed. e-mail: kunihiro@adm.fukuoka-u.ac.jp
© 2016 The Pharmaceutical Society of Japan

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Chem. Pharm. Bull.
Vol. 64, No. 12 (2016)
Chart 1. Synthesis of Symmetrical TAEA-Related Derivatives
Tables S1–S3 for Supplementary materials of the online ver- in 65% yield, and an amide-type C3-symmetrical TAPA de-
sion for details). rivative (21) was also obtained after acylation of TAPA with
Synthesis of targeted CS-symmetrical TAEA derivatives 3,5-ditrifluoromethylbenzoyl chloride (19) in 66% yield (see
(12, 13) was achieved from addition reactions of the starting Experimental).
TAEA (1) to isothiocyanate (6) with different ratios of two re-
All of the structures of the synthesized symmetrical tripo-
agents (1:6=1:2 and 1:6=2:1), and the intermediate adducts dal receptor-type compounds were confirmed by spectroscopic
(10, 11) that were formed were further used for reductive ami- and analytical data. The geometries of the obtained symmetri-
nation with an aldehyde (2d) and NaBH4 in a one-pot reaction cal TAEA and TAPA derivatives described above were con-
to give amine-urea-type TAEA derivatives (12, 13) (see Table firmed from 13C-NMR spectroscopic data14) (see Experimental
S4 for Supplementary materials). The yields of targeted deriv- for details).
atives (12, 13) were about 32–55%. The results indicated that
control of the first step of the addition reaction is not easy.
Biological Activity and Discussion
The high reactivity of isothiocyanate to amine is thought to
In many synthetic receptor-type molecules mimicking the
be the reason for obtaining such experimental results. Amide- supramolecular interaction of biomolecules, it is expected
urea-type TAEA derivatives (14, 15) from the acylations of that the ability to form multiple hydrogen bonds plays an
these isolated compounds (12, 13) with acetic anhydrate (4) important role in guest-molecule recognition. Considering the
were obtained in good to excellent yields (68–92%) (see Table concept called molecular tectonics,15,16) it is also thought that
S5 for Supplementary materials).
a convergent orientation of incorporated functional groups
For the synthesis of some new additional C3-symmetrical is an effective array to produce a multiple hydrogen bond-
tris(3-aminopropyl)amine (TAPA) derivatives, we used the ing interaction between host and guest molecules. From this
same procedure as that described for the preparation of tar- point of view, we prepared title tripodal receptor-type C3- or
geted C3-symmetrical TAEA derivatives. Thus, reductive CS-geometrical molecules (3, 5, 8, 18a, 21) that have donors
amination of TAPA as a starting material with aldehyde (2a) or acceptors for hydrogen bonds on a symmetrical TAEA or
afforded the corresponding amine-type TAPA derivative (18a) TAPA template as new target molecules (Table 1).

Vol. 64, No. 12 (2016)
Chem. Pharm. Bull.
1771
Table 1. Anti-HSV-1 Activity (EC50) and Cytotoxicity (CC50) against Vero Cells of C3-Symmetrical TAEA and TAPA Derivatives
EC50
(µM)
CC50
(µM)
EC50
(µM)
CC50
(µM)
Compound
n
R1
H
R2
Compound
n
R1
H
R2
3a
·H2O
8l
2
>200
>100
20.5
>200
>200
35.9
2
>100
>100
>100
>100
>200
·0.8H2O
3b
2
2
2
H
H
8m
8n
2
2
2
H
H
H
>200
>200
57.3
·0.875CHCl3
3d
·0.5NH3
5a
·0.9H2O
8p
·2-PrOH
Ac
>100
>200
5b
·HCl
·2-PrOH·H2O
2
2
Ac
Ac
>100
>100
>200
>200
20a)
2
3
3
H
H
H
N.D.
>200
>200
17.7
>200
>200
5c
·H2O
18a
·1.2H2O
5d
·0.7H2O
2
2
Ac
Ac
90.7
>200
>200
21
5e
·0.5H2O
>100
a) Data were taken from ref. 6.
Biological evaluations of anti-HSV-1 activities 50% effec- tions of an amine (–NH–) functionality, except for a very low
tive concentration (EC50) by plaque reduction assays17) and level of anti-HSV-1 activity of compound 5d (EC50=90.7 µM),
cytotoxicities against Vero cells 50% cytotoxic concentration also showed no significant anti-HSV-1 (EC50=>100 µM) and
(CC50) of synthesized TAEA and TAPA derivatives (Table 1) cytotoxic (CC50=>200µM) activities.14) Neither of the two
showed that many C3-symmetrical amine-type or urea-type prepared TAPA derivatives 18a and 21, which correspond to
TAEA derivatives (3 or 8) had no significant anti-HSV-1 TAEA derivatives 3a and 20, respectively, showed significant
(EC50=>100 µM) and cytotoxic (CC50=>200µM) activities. anti-HSV-1 (EC50=>100 µM) and cytotoxic (CC50=>200µM)
Among the tested amine-type C3-symmetrical derivatives (3) activities.
shown in Table 1, only compound 3d showed both anti-HSV-1
The most important information from the above-described
(EC50=20.5µM) and cytotoxic (CC50=35.9 µM) activities. Com- results is considered to be the fact that all of the N-acetylated
pound 8p with a large logP value19) (9.94) showed moderate cis–trans mixtures (compounds 5a–d) listed in Table 1
cytotoxic activity (CC50=57.3 µM). Among the C3-symmetrical showed almost no anti-HSV-1 and cytotoxic activities at doses
compounds listed in Table 1 and also the CS-symmetrical de- of <100 µM and <200µM, respectively. Thus, the obtained bio-
rivatives listed in Table 2, there were few distinct correlations logical results indicate at least that the loss of hydrogen atoms
between log P values and anti-HSV-1 activities (EC50 values).
as hydrogen bond donors by N-acetylation of secondary amine
All of the C3-symmetrical amide-type TAEA derivatives (5) moieties in amine-type molecules has a greater influence than
with loss of hydrogen bond donor protons by acylation reac- the geometrical changes of these tripodal receptor-type TAEA

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Chem. Pharm. Bull.
Vol. 64, No. 12 (2016)
Table 2. Anti-HSV-1 Activity (EC50) and Cytotoxicity (CC50) against Vero Cells of CS-Symmetrical TAEA Derivatives
EC50 CC50
(µM) (µM)
EC50 CC50
(µM) (µM)
Compound R1
R2
R3
R4
Compound R1
R2
R3
H
R4
9d
H
13o
H
Ac
>100 250
16.1 21.0
·2H2O
·EtOH
12k
H
14k
H
H
H
H
120.9 >200
>100 >200
>100 57.2
Ac
Ac
H
>100 >200
>100 >200
>100 >200
·0.5H2O
·0.9H2O
12l
H
14l
H
·0.9H2O
·0.5H2O
12o
H
15k
Ac
·0.6H2O
·0.4H2O
13k
H
H
H
16.7 >200
15l
Ac
H
>100 250
1.1 >444
·0.5H2O
13l
H
>100 >200 Aciclovira)
·1.2H2O
a) Data were taken from ref. 18.
derivatives on the expression of anti-HSV-1 activities.
such as 8p and 20, a similar tendency for cytotoxic activity19)
On the other hand, regarding CS-symmetrical molecules was observed (see Table 1). Regarding anti-HSV-1 activities
(as shown in Table 2), many amine-urea-type TAEA deriva- (EC50) of these CS-symmetrical derivatives, amine-urea-type
tives (12k, l, o) that have one urea functionality showed no molecules with N-acetylation of secondary amine moieties
anti-HSV-1 activity (EC50=>100 µM); however, two CS-sym- (see compounds 14k, l, 15k, l) listed in Table 2 also showed
metrical amine-urea-type TAEA derivatives (13k, o) that have almost no anti-HSV-1 activity at a dose of <100 µM, indicat-
two urea functionalities showed considerably high levels of ing that the loss of hydrogen atoms as hydrogen bond donors
anti-HSV-1 activity (EC50=16.7, 16.1 µM, respectively). With by N-acetylation of secondary amine moieties in amine-urea-
regard to the molecules containing an adamantylthiourea type molecules has a greater influence than the geometrical
group(s), we consider that the presence of two adamantylthio- features of these tripodal receptor-type TAEA molecules on
urea groups (13k) is a desirable structural feature in these expression of potential anti-HSV-1 activities.
amine-urea-type derivatives because of almost no anti-HSV-1
As can be seen in the current geometrical and functional
activity of the same geometric compound 12k. Since N- group modifications for the targeted TAEA derivatives [3d
acetylated compounds 14k and 15k, which correspond to CS- (C3)→12k (CS)→13k (CS)→8k (C3)6,20)→14k (CS)→15k (CS)],
symmetrical compounds 12k and 13k, respectively, showed no compound 13k (CS) revealed the maximum magnitude of anti-
anti-HSV-1 activity, the presence of a benzylamino group as HSV-1 activity in these typical examples.
a hydrogen bond donor in the molecule 13k is also required.
On the basis of the obtained structural information on
Compound 13o possessing a high level of cytotoxic activity biological activities in this TAEA series, we are investigating
(CC50=21.0 µM) had large calculated log P values19) over 6.0. further molecular modifications by incorporating a few other
We consider that both the structure and lipophilic property new functional groups that work as hydrogen bonding donors
of the molecule contribute to its overall biological activity, into a TAEA framework in order to confirm the importance
especially its cytotoxic activity. In some C3-type molecules of the presence of hydrogen bonding donors and hopefully to

Vol. 64, No. 12 (2016)
Chem. Pharm. Bull.
1773
find new promising bioactive seeds.
for C30H42N4O6·0.875CHCl3: C, 56.26; H, 6.56; N, 8.50.
Found: C, 56.29; H, 6.62; N, 8.45.
2,2′,2″-[Nitrilotris(2,1-ethanediyliminomethylene)]tris[6-
Experimental
Melting points were determined using a micro melting point methoxyphenol] (3a) (Entry 1) Compound 3a was pre-
apparatus (Yanaco MP-S3) without correction. IR spectra pared from the reaction of 1 with o-vanillin (2a) under the
were measured by a Shimadzu FTIR-8100 IR spectropho- conditions shown in Table S1. Separation of the products
tometer. Low- and high-resolution mass spectra (LR-MS and by centrifugal chromatography (CH2Cl2 :95% EtOH:28%
HR-MS) were obtained by a JEOL JMS HX-110 double-focus- NH3=90:9.5:0.5→85:13:2) gave 3a (1.73g, 62%) as a red-
ing model equipped with an FAB ion source interfaced with a dish purple solid.
l
JEOL JMA-DA 7000 data system. H- and 13C-NMR spectra
3a: mp 35–38°C (mp 42–45°C).15) IR (KBr) cm−1: 3395
were obtained by JEOL JNM A-500 and ECG600R (only for (OH), 3305 (NH), 1590, 1475 (C=C of Ar), 1235, 1075 (C–N
1
compounds 7cm, 9d and 16o). Chemical shifts were expressed and C–O). H-NMR (CDCl3) δ: 2.55 (6H, m, H2′), 2.68 (6H,
l
in δ ppm downfield from an internal TMS signal for H-NMR m, H1′), 3.83 (9H, brs, CH3O–), 3.97 (6H, brs, Hα), 6.27
and the carbon signal of the corresponding solvent [CDCl3 (6H, brs, NH, OH), 6.60 (3H, d, J=7.6Hz, H3), 6.71 (3H, t,
(77.00ppm), dimethyl sulfoxide (DMSO)-d6 (39.50ppm)] for J=7.6Hz, H4), 6.78 (3H, d, J=7.6Hz, H5). 13C-NMR (CDCl3)
13C-NMR. The abbreviations dd=double doublets, dt=double δ: 45.78 (C1′), 51.76 (Cα), 53.90 (C2′), 55.76 (CH3O–), 110.75
triplets, and dm=double multiplets are used for the multiplic- (C5), 118.52 (C4), 120.69 (C3), 122.69 (C2), 147.10 (C1), 147.83
l
ity of H-NMR data. The signal assignments were confirmed (C6). Positive-ion FAB-MS m/z: 555 (M+H)+. HR-FAB-MS
l
by two-dimensional (2D)-NMR analyses: H–lH 2D correlation m/z: 555.3202 (Calcd for C30H43N4O6: 555.3183). Anal. Calcd
l
spectroscopy (COSY), H–l3C heteronuclear multiple-quantum for C30H42N4O6·H2O: C, 62.92; H, 7.74; N, 9.78. Found: C,
l
coherence (HMQC), and H–l3C heteronuclear multiple-bond 62.93; H, 7.71; N, 9.57.
connectivity (HMBC). Microanalyses were performed with a
N2-[(3,4-Dimethoxyphenyl)methyl]-N1,N1-bis[2-[[(3,4-
Yanaco MT-6 CHN corder. Routine monitoring of reactions dimethoxyphenyl)methyl]amino]ethyl]-1,2-ethanediamine
was carried out using precoated Kieselgel 60F254 plates (E. (3c) (Entry 3) Compound 3c was prepared from the re-
Merck). Open and flash column chromatography or centrifu- action of 1 with 3,4-dimethoxybenzaldehyde (2c) under
gal chromatography separations of the reaction products were the conditions shown in Table S1. Separation of the prod-
performed on silica gel (Kanto 60N or Able-Biott) with a UV ucts by flash chromatography (CH2Cl2 :95% EtOH:28%
detector. Commercially available starting materials were used NH3=93:6.6:0.4→85:14:1) gave 3c (5.00g, 84%) as a sticky
without further purification, and dry solvents were used in all yellow oil.
1
reactions.
3c: H-NMR (CDCl3) δ: 2.75–2.77 (12H, m, H1′, H2′), 3.84
General Procedure for the Preparation of C3-Symmetri- (6H, s, Hα), 3.85 (9H, s, OCH3 on C3), 3.90 (9H, s, OCH3 on
cal Amine-Type TAEA Derivatives (3): Example: Synthesis C4), 4.36 (3H, brs, NH), 6.79 (3H, d, J=8.2Hz, H5′), 6.86 (3H,
of 4,4′,4″-[Nitrilotris(2,1-ethanediyliminomethylene)]tris(2- dd, J=8.2, 2.1Hz, H6′), 7.13 (3H, d, J=2.1Hz, H2′). 13C-NMR
methoxyphenol) (3b) (Entry 2) (Step 1) To a solution of (CDCl3) δ: 45.21 (C1′), 52.24 (Cα), 53.02 (C2′), 55.92 (OCH3
vanillin (2b, 2.28g, 15.0mmol) in methanol (MeOH, 6mL) on C4), 56.22 (OCH3 on C3), 111.16 (C5), 112.58 (C2), 121.45
was added TAEA (1, 731mg, 5.00mmol) in MeOH (3mL) at (C6), 127.88 (C1), 149.02 (C3), 149.39 (C4). Positive-ion FAB-
room temperature (r.t.) under an N2 atmosphere. After stirring MS m/z: 597 (M+H+). HR-FAB-MS m/z: 597.3660 (Calcd for
for 1h, the yellow solution changed to a yellow sticky suspen- C33H49N4O6: 597.3652). Anal. Calcd for C33H48N4O6·2.4H2O: C,
sion.
(Step 2) After dilution with MeOH (12mL) and ice-cool-
61.93; H, 8.32; N, 8.45. Found: C, 61.99; H, 8.17; N, 8.72.
N2-(1,3-Benzodioxol-5-ylmethyl)-N1,N1-bis[2-[(1,3-
ing, compound 1 (1.84g, 10.0mmol) in dry benzene (20mL) benzodioxol-5-ylmethyl)amino]ethyl]-1,2-ethanediamine
was added dropwise with stirring. After stirring for 20min at (3d) (Entry 4) Compound 3d was prepared from the reac-
r.t., the reaction mixture was refluxed for 1h. After cooling to tion of 1 (1.46g, 10.0mmol) with piperonal (2d) under the
r.t., a small amount of NaBH4 (945mg, 25.0mmol) was added conditions shown in Table S1. Separation of the products
to the mixture and the mixture was stirred for 2.5h at 0°C. It by centrifugal chromatography (CH2Cl2 :95% EtOH:28%
was then diluted with aqueous ammonium chloride (NH4Cl, NH3=85:14:1) gave 3d (0.760g, 14%) as a pale yellow oil.
10%, 100mL) and extracted with CH2Cl2 (3×200mL). The
(Entry 5) This compound was also prepared as follows:
combined organic layer was washed with brine (50mL) and To a solution of 2d (2.25g, 15.0mmol) in benzene (40mL)
dried over anhydrous magnesium sulfate (MgSO4). Evapora- were added compound 1 (731mg, 5.0mmol) and p-toluene-
tion of the solvent gave crude 3b (2.43g, 88%). An analytical sufonic acid (TsOH, 66mg, 0.35mmol) and the resulting
sample of 3b was obtained by recrystallization from chloro- mixture was refluxed. The resultant H2O was removed with a
form (CHCl3) as an off-white solid.
Dean–Stark distillation apparatus for 18h under an N2 atmo-
3b: mp 64–69°C (from CHCl3). IR (KBr) cm−1: 1279, 1034, sphere. After evaporation of the solvent, the resulting mixture
1
1125 (C–O). H-NMR (DMSO-d6) δ: 2.5–2.7 (12H, m, H1′, was diluted with MeOH (30mL) and then a small amount of
2′), 3.21 (6H, brs, NH, OH), 3.54 (6H, brs, Hα), 3.71 (9H, brs, NaBH4 (1.57mg, 41.5mmol) was added and the mixture was
OCH3), 6.60–6.73 (6H, m, H5, 6), 6.84 (3H, d, J=1.5Hz, H3), stirred for 3h at 0°C under an N2 atmosphere. After evapo-
8.30 (<1H, s, CHCl3). 13C-NMR (DMSO-d6) δ: 46.56 (C1′), ration of the solvent, CHCl3 (20mL), brine (5mL), and 5%
52.75 (Cα), 54.00 (C2′), 55.44 (OCH3), 79.08 (CHCl3), 112.07 NaHCO3 (5mL) were added to the obtained yellow solid, and
(C3), 114.99 (C6), 120.14 (C5), 131.60 (C4), 145.09 (C1), 147.31 then the resulting mixture was vigorously stirred for 1h. After
(C2). Positive-ion FAB-MS m/z: 555 (M+H+). HR-FAB-MS separation of the organic layer, the aqueous layer was re-
m/z: 555.3185 (Calcd for C30H43N4O6: 555.3183). Anal. Calcd extracted with CHCl3 (2×20mL). The combined organic layer

1774
Chem. Pharm. Bull.
Vol. 64, No. 12 (2016)
was dried over MgSO4, and evaporation of the solvent gave a 149.60 (C4), 170.87 (C=O). (Signals of only the predominant
brown oil. The product was purified by flash chromatography conformer were assigned.) Positive-ion FAB-MS m/z: 723
(CH2Cl2 :95% EtOH:28% NH3=73:25:2→65:32:3) to give (M+H+). HR-FAB-MS m/z: 723.3967 (Calcd for C39H55N4O9:
3d (1.38g, 50%) as a yellow oil. An analytical sample of 3d 723.3969). Anal. Calcd for C39H54N4O9·H2O: C, 63.22; H, 7.62;
was obtained as a pale yellow solid by centrifugal chromatog- N, 7.56. Found: C, 63.18; H, 7.70; N, 7.61.
raphy (CH2Cl2 : 95% EtOH : 28% NH3=80:19.5:0.5).
N,N′,N″-[2,2′,2″-Nitrilotris(ethane-2,1-diyl)]tris[N-(2-hy-
3d: mp 215°C (dec.). IR (NaCl) cm−1: 3300 (NH of amine), droxy-3-methoxybenzyl)acetamide] (5a) (Entry 7) Com-
2825 (CH2 of methylenedioxy), 1245, 1040 (=C–O–C–), 930 pound 5a was prepared by acylation of 3a with Ac2O (4)
1
(C–O). H-NMR (DMSO-d6) δ: 2.76 (6H, t, J=5.2Hz, H2′), under the conditions shown in Table S2. After work-up, crude
3.06 (6H, t, J=5.2Hz, H1′), 4.11 (6H, s, Hα), 6.04 (6H, s, H2), compound 5a (1.59g, 75%) was obtained as a white solid. Re-
6.93 (3H, d, J=7.9Hz, H7), 7.07 (3H, dd, J=7.9, 1.5Hz, H6), crystallization from AcOEt gave an analytically pure product
7.27 (3H, d, J=1.5Hz, H4), 9.48 (3H, brs, NH). 13C-NMR 5a as colorless crystals.
(DMSO-d6) δ: 43.28 (C1′), 49.54 (C2′ or Cα), 49.62 (Cα or
5a: mp 107–109°C (from AcOEt). IR (KBr) cm−1: 3150 (OH),
1
C2′), 101.16 (C2), 108.13 (C7), 110.42 (C4), 124.24 (C6), 125.14 1625 (C=O), 1070 (C–O of phenol, ether). H-NMR (DMSO-
1
(C5), 147.22 (C3a), 147.58 (C7a). H-NMR (CDCl3) δ: 2.17 (3H, d6) δ: 1.98, 1.99, 2.02, 2.03* (9H, s, CH3CO–), 2.36–3.47 (3H,
brs, NH), 2.56 (6H, t, J=5.8Hz, H2′), 2.64 (6H, t, J=5.8Hz, m, H2′), 2.50–2.58 (3H, m, H2′), 3.13–3.24 (6H, m, H1′),
H1′), 3.64 (6H, s, Hα), 5.90 (6H, s, H2), 6.70 (6H, brs, H6, 7), 3.74*, 3.78 (9H, s, CH3O–), 4.38, 4.40, 4.42* (6H, brs, Hα),
6.79 (3H, s, H4). 13C-NMR (CDCl3) δ: 46.88 (C1′), 53.57 (Cα), 6.57–6.76 (6H, m, H3, 4), 6.83–6.88 (3H, m, H5), 8.76, 8.79,
54.29 (C2′), 100.76 (C2), 107.95 (C7), 108.55 (C4), 121.08 (C6), 8.82, 8.95, 8.96*, 8.98 (3H, brs, OH). (The signals of the
134.14 (C5), 146.41 (C7a), 147.62 (C3a). Positive-ion FAB-MS predominant conformer were asterisked.) 13C-NMR (DMSO-
m/z: 549 (M+H)+. HR-FAB-MS m/z: 549.2728 (Calcd for d6) δ: 20.64, 20.86, 20.90*, 20.93, 21.28, 21.32* (CH3CO–),
C30H37N4O6: 549.2713). Anal. Calcd for C30H36N4O6·0.5NH3: 43.05, 43.31, 43.37, 43.52, 43.77 (Cα, C1′), 46.16, 46.24*, 46.40
C, 64.67; H, 6.78; N, 11.31. Found: C, 64.42; H, 6.81; N, 11.04. (C1′), 47.39, 47.51* (Cα), 51.07, 51.32*, 51.83, 52.07* (C2′),
N1-(4-Pyridinylmethyl)-N2,N2-bis[2-[(4-pyridinylmethyl)- 55.70*, 55.79* (CH3O–), 110.98, 111.01, 111.10, 111.17*, 111.27
amino]ethyl]-1,2-ethanediamine (3e) (Entry 6) Compound (C5), 118.61, 118.82*, 119.51 (C4), 119.61, 119.79, 121.25* (C3),
3e was prepared from the reaction of 1 with isonicotinalde- 123.78, 123.86, 124.09* (C2), 143.99, 144.06, 144.50* (C1),
hyde (2e) under the conditions shown in Table S1. Separation 147.38*, 147.61, 147.64, 147.68 (C6), 169.78, 169.84, 170.43,
of the products by centrifugal chromatography (CH2Cl2 :95% 170.54*, 170.67 (C=O). (Signals of only the predominant
EtOH:28% NH3=73:15:2→65:32:3) gave 3e (3.97g, 95%) conformer were asterisked.) Positive-ion FAB-MS m/z: 681
as a yellow oil.
(M+H)+. HR-FAB-MS m/z : 681.3517 (Calcd for C36H49N4O9:
1
3e: H-NMR (CDCl3) δ: 2.00 (3H, brs, NH), 2.61 (6H, t, 681.3500). Anal. Calcd for C36H48N4O9·0.9 H2O: C, 62.04; H,
J=5.2Hz, H2′), 2.67 (6H, t, J=5.2Hz, H1′), 3.76 (6H, s, Hα), 7.20; N, 8.04. Found: C, 62.09; H, 7.15; N, 7.85.
7.20 (6H, dd, J=4.6, 1.5Hz, H3, 5), 8.50 (6H, dd, J=4.6,
N,N′,N″-(Nitrilotri-2,1-ethanediyl)tris[N-(4-hydroxy-
1.5Hz, H2, 6). 13C-NMR (CDCl3) δ: 47.17 (C1′), 52.73 (Cα), 3-methoxyphenylmethyl)acetamide] Hydrochloride (5b·HCl)
54.20 (C2′), 122.78 (C3, 5), 149.33 (C4), 149.80 (C2, 6). and [[(Nitrilotri-2,1-Ethanediyl)tris(acetylazanediyl)]tris-
KM4-4-1 Positive-ion FAB-MS m/z: 420(M+H+). HR-FAB- (methylene)]tris(2-methoxybenzene-4,1-diyl)
MS m/z: 420.2873 (Calcd for C24H34N7: 420.2876).
Triacetate
(5b′) (Entry 8) Compound 5b′ was prepared by acylation
General Procedure for the Preparation of C3-Symmetri- of compound 3b (0.943g, 1.70mmol) with Ac2O (4, 1.39g,
cal Amide-Type TAEA Derivatives (5): Example: Synthe- 13.6mmol) under the conditions shown in Table S2. After
sis of N,N′,N″-(Nitrilotri-2,1-ethanediyl)tris[N-(3,4-dime- work-up crude compound 5b′ (987mg, 72%) was obtained as
thoxyphenylmethyl)acetamide] (5c) (Entry 9) To a solution a white solid. After dilution of this material with EtOH (4mL)
of compound 3c (2.85g, 5.00mmol) in benzene (8mL) was and addition of 1M HCl (in EtOH), evaporation of the solvent
added dropwise a solution of acetic anhydride (Ac2O, 4, 1.55g, and azeotropic evaporation with dry benzene were repeated.
15.0mmol) in benzene (2mL) at r.t., and then the mixture was Then the white semisolid material was recrystallized from
stirred for 2h. After removal of the solvent by evaporation, 1M 2-PrOH to give an analytically pure salt of the deacylated tar-
NaOH solution (20mL) was added to the resulting residue and get compound 5b·HCl as colorless crystals.
then the mixture was extracted with EtOAc (3×40mL). The
5b·HCl: mp 69–73°C (from 2-PrOH). IR (KBr) cm−1: ca.
organic layer was washed with brine (25mL) and dried with 3200 (OH), ca. 2600 (NH+), 1636 (C=O), 1279, 1032, 1125
1
MgSO4 and then evaporated to give a pale yellow oil, which (C–O). H-NMR (DMSO-d6) δ: 1.04 (6H, d, J=6.1Hz, CH3
was purified by flash chromatography (EtOH:EtOAc=4:6) to of 2-PrOH), 2.10*, 2.11 (9H, s, COCH3), 3.17, 3.24*, 3.33
give 5c (2.88g, 80%) as a colorless semisolid.
(6H, brs, H2′), 3.39 (4H, brs, OH) 3.66*, 3.75 (6H, brs, H1′),
5c: mp 34–39°C. IR (KBr) cm−1: 1646 (C=O), 1261, 1233 3.75, 3.76, 3.77*, 3.78 (9H, s, OCH3), 3.80 (1H, q, J=6.1Hz,
(C–O of aromatic ether), 1137, 1025 (C–O of aliphatic ether). CH of 2-PrOH), 4.38, 4.48* (9H, brs, Hα), 6.65* (0.8H, dd,
1H-NMR (CDCl3) δ: 2.07, 2.09*, 2.11, 2.15 (9H, s, COCH3), J=7.9, 1.8Hz, H5), 6.68–6.73 (0.4H, m, H5, 6), 6.79* (0.8H,
2.05, 2.52, 2.58*, 2.63 (6H, t, J=7.3Hz, H2′), 3.17, 3.23, 3.34*, d, J=7.9Hz, H6), 6.82* (0.8H, brs, H3), 6.88–6.93 (0.2H, m,
3.37 (6H, t, J=7.3Hz, H1′), 3.83, 3.84, 3.86, 3.86* (18H, s, H3), 8.96*, 9.85, 10.88, 11.73 (3H, brs, NH, OH). (The signals
OCH3), 4.39, 4.43*, 4.47, 4.50 (6H, s, Hα), 6.63–6.79 (6H, m, of the predominant conformer were asterisked.) 13C-NMR
H2, 6), 6.80–6.86 (3H, m, H5). (The signals of the predomi- (DMSO-d6) δ: 21.40 (COCH3), 25.39 (CH3 of 2-PrOH), ca.
nant conformer were asterisked.) 13C-NMR (CDCl3) δ: 21.64 40 (C1′), 50.33 (C2′), 51.29 (Cα), 55.64 (OCH3), 61.93 (CH of
(COCH3), 44.28 (C1′), 52.03 (C2′), 52.50 (Cα), 55.98 (OCH3), 2-PrOH), 111.52 (C3), 115.64 (C6), 119.23 (C4), 127.56 (C5),
109.76 (C2), 111.61 (C5), 118.51 (C6), 129.12 (C1), 148.74 (C3), 146.01 (C1), 147.77 (C2), 171.38 (C=O). (Signals of only the

Vol. 64, No. 12 (2016)
Chem. Pharm. Bull.
1775
predominant conformer were assigned.) Positive-ion FAB- yellow sticky oil.
MS m/z: 681 (M+H+). HR-FAB-MS m/z: 681.3491 (Calcd for
5e: IR (KBr) cm−1: 1636 (C=O), 796 (CH of pyridine).
C36H49N4O9: 681.3500). Anal. Calcd for C36H48N4O9·HCl·2- 1H-NMR (CDCl3) δ: 2.02, 2.04*, 2.07, 2.17, 2.21 (9H, s, CH3),
PrOH·H2O: C, 58.89; H, 7.48; N, 7.04. Found: C, 58.75; H, 2.50–2.70 (6H, m, H2′), 3.18–3.45 (6H, m, H1′), 4.45, 4.50*,
7.70; N, 6.84.
4.55, 4.56, 4.58 (6H, s, Hα), 7.05–7.17 (6H, m, H3, 5), 8.52,
5b′: Positive-ion FAB-MS m/z: 807 (M+H+). HR-FAB-MS 8.60* (6H, t, J=6.1Hz, H2, 6). (The signals of the predomi-
1
m/z: 807.3812 (Calcd for C42H55N4O12: 807.3816). Both H- and nant conformer were asterisked.) 13C-NMR (CDCl3) δ: 21.46
13C-NMR in CDCl3 were too complicated and we could not (CH3), 44.55 (C1′), 51.59 (C2′), 51.78 (Cα), 121.15 (C3, 5),
provide full assignments of the observed signals.
145.92 (C4), 150.41 (C2, 6), 170.94 (C=O). (Signals of only
N,N',N″-(Nitrilotri-2,1-ethanediyl)tris[N-(1,3-benzo- the predominant conformer were assigned.) Positive-ion FAB-
dioxol-5-ylmethyl)acetamide] (5d) and N,N′-[[[2-[(1,3- MS m/z: 546 (M+H+). HR-FAB-MS m/z: 546.3194 (Calcd for
Benzodioxol-5-ylmethyl)amino]ethyl]imino]di-2,1-eth- C30H40N7O3: 546.3193). Anal. Calcd for C30H39N7O3·0.5H2O:
anediyl]bis[N-(1,3-benzodioxol-5-ylmethyl)acetamide] (9d) C, 64.96; H, 7.27; N, 17.68. Found: C, 65.00; H, 7.20; N, 17.46.
(Entry 10) Compound 5d was prepared by acylation of 3d
General Procedure for the Preparation of C3-Sym-
with Ac2O (4) under the conditions shown in Table S2. Sepa- metrical Urea-Type TAEA Derivatives (7, 8): Example:
ration of the products by flash chromatography (CH2Cl2 :95% Synthesis of N,N″,N-(Nitrilotri-2,1-ethanediyl)tris[N′-(3,4-
EtOH:28% NH3=95:4.7:0.3→93:6.6:0.4) gave 5d (2.67g, dimethoxyphenyl)thiourea] (8l) (Entry 14) To a solution
53%) as a colorless semisolid and 9d (0.94g, 20%) as a color- of 3,4-dimethoxyphenyl isothiocyanate (6l, 2.93g, 15.0mmol)
less oil. Compound 9d was purified by centrifugal chromatog- in CH2Cl2 (20mL) was added TAEA (1, 731mg, 5.00mmol) at
raphy (EtOAc:EtOH=99.5:0.5) to give an analytically pure r.t. After stirring for 1h, the resulting white solid was filtrated
9d as a hygroscopic white semisolid.
to obtain crude compound 8l (3.35g, 91%). Recrystallization
5d: mp 46–65°C. IR (KBr) cm−1: 1644 (C=O), 1248, 1037 from MeOH gave analytically pure compound 8l as a white
1
(=C–O–C–), 923 (C–O of methylenedioxy). H-NMR (CDCl3) solid.
δ: 2.09*, 2.10, 2.11, 2.14 (9H, s, CH3), 2.40–2.70 [6H, 2.43
8l: mp 161–163°C (from MeOH). IR (KBr) cm−1: 1513
1
(brs), 2.49, 2.55* (t, J=7.0Hz), 2.65 (brs), H2′], 3.10–3.45 (C=S), 1258, 1235, 1134, 1027 (C–O). H-NMR (CDCl3) δ:
[6H, 3.16, 3.22, 3.31* (t, J=7.0Hz), 3.37 (brs), H1′], 4.36, 2.67 (6H, t, J=6.9Hz, H2′), 3.48–3.55 (6H, m, H1′), 3.72 (9H,
4.39*, 4.44, 4.46 (6H, s, Hα), 5.91, 5.91, 5.94*, 5.95, 5.96 (6H, s, –OCH3), 3.73 (9H, s, –OCH3), 6.79 (3H, dd, J=8.5, 2.1Hz,
s, H2), 6.57–6.80 (9H, m, ArH). (The signals of the predomi- H6), 6.90 (3H, d, J=8.5Hz, H5), 6.96 (3H, d, J=2.1Hz, H2),
nant conformer were asterisked.) 13C-NMR (CDCl3) δ: 21.54 7.36 (3H, brs, Hβ), 9.37 (3H, brs, Hα). 13C-NMR (CDCl3)
(CH3), 44.06 (C1′), 51.84 (C2′), 52.50 (Cα), 101.15 (C2), 106.79 δ: 41.97 (C1′), 52.19 (C2′), 55.45 (–OCH3), 55.70 (–OCH3),
(C4), 108.45 (C7), 119.57 (C6), 130.41 (C5), 147.14 (C3a or 109.23 (C2), 111.97 (C5), 116.28 (C6), 131.62 (C1), 146.27 (C4),
7a), 148.27 (C7a or 3a), 170.71 (C=O). (Signals of only the 148.64 (C3), 180.28 (C=S). Positive-ion FAB-MS m/z: 732
predominant conformer were assigned.) Positive-ion FAB- (M+H+). HR-FAB-MS m/z: 723.2682 (Calcd for C33H46N7O6S3:
MS m/z: 675 (M+H+). HR-FAB-MS m/z: 675.3027 (Calcd for 732.2672). Anal. Calcd for C33H45N7O6S3·0.8H2O: C, 53.10; H,
C36H43N4O9: 675.3030). Anal. Calcd for C36H42N4O9·0.7H2O: 6.29 ; N, 13.14. Found: C, 53.08; H, 6.09; N, 13.22.
C, 62.91; H, 6.36 ; N, 8.15. Found: C, 62.90; H, 6.57; N, 8.01.
N, N ″, N ⁗-(Nitrilotri-2,1-ethanediyl)tris[N-(3,4-
9d: mp 76–81°C. IR (KBr) cm−1: 1638(C=O), 1251, dimethoxybenzyl)-N′-[3-(trifluoromethyl)phenyl]urea]
1
1037 (=C–O–C–), 924 (C–O of methylenedioxy). H-NMR (7cm) (Entry 12) Compound 7cm was prepared from 3c
(DMSO-d6) δ: 1.44 (1H, brs, NH), 1.98*, 1.99 (5H, s, CH3), (282mg, 0.473mmol) and 3-(trifluoromethyl)phenyl isocya-
2.02 (0.6H, s, CH3), 2.06 (0.4H, s, CH3), 2.29–2.47 (4H, nate (6m, 266mg, 1.41mmol) under the conditions shown in
m, H2′), 2.50–2.57 (1H, m, H2″), 2.61–2.69 (1H, m, H2″), Table S3. Separation of the product by flash chromatography
3.05–3.31 (6H, m, H1′, 1‴), 3.45–5.57 (1H, m, Hα′), 3.83–3.89 (CH2Cl2 :95% EtOH:28% NH3=97:2.7:0.3→95:4.7:0.3) gave
(1H, m, Hα′), 4.32*, 4.36, 4.39, 4.44 (4H, s, Hα), 5.98*, 5.99 7cm (242mg, 44%) as a colorless oil.
1
(4H, s, H2), 6.01*, 6.05 (2H, s, H2‴), 6.62–6.70 (2H, m,
7cm: H-NMR (CDCl3) δ: 1.64 (3H, s, NH), 2.79 (6H, brt,
H6), 6.73–6.77 (2H, m, H4), 6.79–6.92 (4H, m, H7, 6‴, 7‴), J=6.6Hz, H2′), 3.55 (6H, brs, H1′), 3.82 (9H, s, MeO on C3),
7.03–7.08 (1H, m, H4‴). 13C-NMR (DMSO-d6) δ: 21.50 (CH3), 3.84 (9H, s, MeO on C4), 4.74 (6H, s, Hα), 6.75–6.80 (9H, m,
43.30 (C1″), 45.57 (C1′), 47.55 (Cα), 49.99 (C2″), 51.56 (C2′), H2, 5, 6), 7.19 (3H, brd, J=7.5Hz, H4″), 7.23 (3H, t, J=7.5Hz,
58.48 (Cα′), 100.91 (C2‴), 101.09 (C2), 108.06 (C4), 108.14 H5″), 7.36 (3H, brd, J=7.5Hz, H6″), 7.58 (3H, brs, H2″).
(C7‴ or C7), 108.36 (C7 or C7‴), 110.18 (C4‴), 119.79 (C6), 13C-NMR (CDCl3) δ: 45.44 (C1′), 51.03 (Cα), 52.68 (C2′),
123.82 (C6‴), 128.34 (C5‴), 132.05 (C5), 146.32 (C7a), 147.05 55.89 (CH3O), 55.92 (CH3O), 110.24 (C2), 111.31 (C5), 116.40
(C7a‴), 147.26 (C3a‴), 147.72 (C3a), 169.78 (C=O). (Signals of (q, J=4.3Hz, C2″), 119.28 (q, J=4.3Hz, C4″), 122.83 (C6″),
only the predominant conformer were assigned.) Positive-ion 123.89 (q, J=271.6Hz, CF3), 129.07 (C1), 129.19 (C5″), 131.02
FAB-MS m/z: 633 (M+H+). HR-FAB-MS m/z: 633.2918 (Calcd (q, J=31.8Hz, C3″), 139.72 (C1″), 148.92 (C4), 149.66 (C3),
for C34H41N4O8: 633.2924). Anal. Calcd for C34H40N4O8·2H2O: 155.87 (C=O). Positive-ion FAB-MS m/z: 1158 (M+H+). HR-
C, 61.07; H, 6.63 ;N, 8.38. Found: C, 61.01; H, 6.76; N, 8.13.
N,N',N″-(Nitrilotri-2,1-ethanediyl)tris[N-(pyridin-4-yl-
methyl)acetamide] (5e) (Entry 11) Compound 5e was ylmethyl)-N′-[4-(trifluoromethyl)phenyl]thiourea]
FAB-MS m/z: 1158.4369 (Calcd for C57H61F9N7O9: 1158.4387).
N,N″,N⁗-(Nitrilotri-2,1-ethanediyl)tris[N-(pyridin-4-
(7ep)
prepared by acylation of compound 3e with Ac2O (4) under (Entry 13) Compound 7ep was prepared from 3e (242mg,
the conditions shown in Table S2. Separation of the prod- 0.577mmol) and 4-(trifluoromethyl)phenyl isothiocyanate (6p,
ucts by flash chromatography (CH2Cl2 :95% EtOH:28% 352mg, 1.73mmol) under the conditions shown in Table S3.
NH3=90:9.5:0.5→85:14:1) gave 5e (2.51g, 61%) as a pale After evaporation of the solvent, recrystallization of the result-

1776
Chem. Pharm. Bull.
Vol. 64, No. 12 (2016)
ing mixture from 2-PrOH gave 7ep (296mg, 50%) as a white J=8.4Hz, H2, 6), 7.93 (3H, brs, NHβ), 9.88 (3H, s, NHα).
solid.
13C-NMR (DMSO-d6) δ: 41.81 (C1′), 51.86 (C2′), 121.77
1
7ep: mp 122–127°C (from 2-PrOH). H-NMR (DMSO-d6) δ: (C2, 6), 123.34 (q, J=32.1Hz, C4), 124.28 (q, J=271.0Hz,
2.95 (6H, t, J=6.7Hz, H2′), 3.88 (6H, brs, H1′), 5.10 (6H, brs, CF3), 125.50 (q, J=3.1Hz, C3, 5), 143.19 (C1), 180.22 (C=S).
Hα), 7.18 (6H, d, J=5.6Hz, H3, 5), 7.48 (6H, d, J=8.4Hz, H2″, Positive-ion FAB-MS m/z: 756 (M+H+). HR-FAB-MS m/z:
6″), 7.59 (6H, d, J=8.4Hz, H3″, 5″), 8.50 (6H, d, J=5.6Hz, H2, 756.1658 (Calcd for C30H31F9N7S3: 756.1659). Anal. Calcd for
6), 9.84 (3H, brs, NH). 13C-NMR (DMSO-d6) δ: 49.65 (C1′), C30H30F9N7S3·2-PrOH : C, 48.58; H, 4.69; N, 12.02. Found: C,
51.25 (C2′), 53.10 (Cα), 121.69 (C3, 5), 124.40 (q, J=32.1Hz, 48.54; H, 4.59; N, 11.96.
C4″), 124.74 (q, J=271.1Hz, CF3), 124.94 (q, J=4.1Hz, C3″,
General Procedure for the Preparation of CS-Sym-
5″), 125.23 (C2″, 6″), 144.35 (C1″), 145.99 (C4), 149.56 (C2, 6), metrical Amine-Urea-Type TAEA Derivatives (12, 13):
182.21 (C=S). Positive-ion FAB-MS m/z: 1029 (M+H+). HR- Example: Synthesis of N-[2-[Bis[2-[(1,3-benzodioxol-5-yl-
FAB-MS m/z: 1029.2927 (Calcd for C48H46F9N10S3: 1029.2925). methyl)amino]ethyl]amino]ethyl]-N′-(tricyclo[3.3.1.13,7]-
N,N″,N-(Nitrilotri-2,1-ethanediyl)tris[N′-[3-(trifluoro- dec-1-yl)thiourea (12k) (Entry 18) (Step 1) To a solution
methyl)phenyl]urea] (8m) (Entry 15) Compound 8m was of TAEA (1, 1.46g, 10.0mmol) in CH2Cl2 (160mL) was added
prepared from 1 (292mg, 2.00mmol) and 3-(trifluoromethyl)- 1-adamantyl isothiocyanate (6k, 2.93g, 15.0mmol) in CH2Cl2
phenyl isocyanate (6m, 1.12g, 6.00mmol) under the conditions (20mL) and the mixture was stirred for 1h at r.t. Evaporation
shown in Table S3. After filtration of the resulting precipitates, of the solvent gave a pale yellow oil.
recrystallization from propionitrile (EtCN) gave 8m (1.19g,
(Step 2) To a solution of the resulting oil in MeOH (5mL)
was added a solution of piperonal (2d, 3.00g, 20.0mmol) in
84%) as colorless crystals.
8m: mp 187–190°C (from EtCN). IR (KBr) cm−1: 3334 MeOH (10mL) at r.t. under an argon atmosphere and the mix-
1
(NH), 1645 (CONH), 1340 (CF3). H-NMR (DMSO-d6) δ: 2.63 ture was stirred for 20h.
(6H, t, J=6.7Hz, H2′), 3.17–3.25 (6H, m, H1′), 6.25 (3H, t,
(Step 3) To the resulting yellow solution was added
J=5.5Hz, NHβ), 7.20 (3H, dd, J=7.6, 0.9Hz, H4), 7.41 (3H, t, MeOH (80mL) and NaBH4 (2.04g, 54.0mmol) at 0°C under
J=7.9Hz, H5), 7.49 (3H, d, J=8.5, H6), 7.93 (3H, s, H2), 8.87 an argon atmosphere with stirring for 4h, and then stir-
(3H, s, NHα). 13C-NMR (DMSO-d6) δ: 37.53 (C1′), 53.72 (C2′), ring was continued at r.t. for 15h. After evaporation of the
113.54 (q, J=4.1Hz, C2), 117.08 (q, J=4.1Hz, C4), 121.06 solvent, aqueous ammonium acetate (NH4OAc, 10%) was
(C6), 124.16 (q, J=272.1Hz, CF3), 129.34 (q, J=31.0, C3), added to the resulting white solid and the mixture was ex-
129.52 (C5), 141.24 (C1), 155.03 (C=O). Positive-ion FAB- tracted with CHCl3 (3×200mL). The separated organic layer
MS m/z: 708 (M+H+). HR-FAB-MS m/z: 708.2352 (Calcd for was dried over MgSO4 and filtrated, and evaporation of
C30H31F9N7O3: 708.2345). Anal. Calcd for C30H30F9N7O3: C, the solvent gave a pale yellow oil. Separation of the prod-
50.92; H, 4.27; N, 13.86. Found: C, 50.84; H, 4.07; N, 13.89.
ucts by flash chromatography (CH2Cl2 :95% EtOH:28%
N,N″,N-(Nitrilotri-2,1-ethanediyl)tris[N′-[4-(trifluoro- NH3=950:47:3→73:25:2) gave 16k (403mg, 17%) as a white
methyl)phenyl]urea (8n) (Entry 16) Compound 8n was solid, 13k (365mg, 6%) as a white solid, and 12k (2.03g, 33%)
prepared from 1 (292mg, 2.00mmol) and 4-(trifluoromethyl)- as a colorless semisolid.
phenyl isocyanate (6n, 1.12g, 6.00mmol) under the conditions
12k: mp 46–52°C. IR (KBr) cm−1: 3275 (NH of sec-amine),
shown in Table S3. After filtration of the resulting precipitates, 1489 (C=S), 1247, 1038, 929 (C–O). 1H-NMR (CDCl3) δ:
recrystallization from EtOH gave 8n (1.24g, 88%) as colorless 1.58–1.67 (6H, m, Ha), 1.99 (6H, brs, Hb), 2.02 (3H, brs, Hc),
crystals.
2.35 (3H, brs, NH), 2.61 (4H, brt, J=6.1Hz, H2′), 2.66–2.71
8n: mp 237–238°C (from EtOH). IR (KBr) cm−1: 1649 (6H, m, H1′, 2″), 3.67 (4H, s, Hα), 5.92 (4H, s, H2), 6.09 (1H,
(CONH), 1329 (CF3). H-NMR (DMSO-d6) δ: 2.63 (6H, t, brs, NH), 6.75 (4H, brs, H4, 6), 6.81 (2H, s, H7). 13C-NMR
1
J=6.4Hz, H2′), 3.22 (6H, dt, J=6.4, 6.1Hz, H1′), 6.28 (3H, (CDCl3) δ: 29.44 (Cc), 36.11 (Ca), 42.03 (Cb), 43.06 (C1″),
t, J=5.5Hz, NHβ), 7.52 (6H, d, J=8.9Hz, H3, 5), 7.56 (6H, d, 46.63 (C1′), 53.43, 53.55, 53.62, 53.72 (C2′, 2″, α, d), 100.91
J=8.9Hz, H2, 6), 8.91 (3H, s, NHα). 13C-NMR (DMSO-d6) δ: (C2), 108.14 (C4), 108.67 (C7), 121.37 (C6), 133.34 (C5),
37.52 (C1′), 53.65 (C2′), 117.20 (C2, 6), 120.89 (q, J=32.1Hz, 146.72 (C3a), 147.78 (C7a), 180.73 (C=S). Positive-ion FAB-
C4), 124.54 (q, J=271.0Hz, CF3), 125.74 (q, J=4.1Hz, C3, 5), MS m/z: 608 (M+H+). HR-FAB-MS m/z: 608.3273 (Calcd for
144.09 (C1), 154.84 (C=O). Positive-ion FAB-MS m/z: 708 C33H46N5O4S: 608.3271). Anal. Calcd for C33H45N5O4S·0.5H2O:
(M+H+). HR-FAB-MS m/z: 708.2352 (Calcd for C30H31F9N7O3: C, 64.26; H, 7.52; N, 11.35. Found: C, 64.32; H, 7.64; N, 11.25.
1
708.2345). Anal. Calcd for C30H30F9N7O3: C, 50.92; H, 4.27; N,
13.86. Found: C, 50.88; H, 4.20; N, 13.92.
16k: H-NMR (CDCl3) δ: 1.69 (6H, brs, Ha), 2.13 (3H,
brs, Hc), 2.15 (6H, brs, Hb), 2.70 (2H, dt, J=5.5, 0.3Hz, H5),
N,N″,N-(Nitrilotri-2,1-ethanediyl)tris[N′-[4-(trifluoro- 3.62–3.68 (2H, m, H4), 6.49 (0.5H, brs, NH), 7.28 (0.5H, brs,
methyl)phenyl]-thiourea] (8p) (Entry 17) Compound NH). 13C-NMR (CDCl3) δ: 29.54 (Cc), 36.19 (Ca), 42.11 (Cb),
8p was prepared from 1 (292mg, 2.00mmol) and 4- 43.01 (C4), 53.53 (C5), 54.25 (Cd), 180.49 (C=S). Positive-ion
(trifluoromethyl)phenyl isothiocyanate (6p, 1.12g, 6.00mmol) FAB-MS m/z: 237 (32, M+H+). HR-FAB-MS m/z: 237.1417
under the conditions shown in Table S3. Filtration of the re- (Calcd for C13H21N2S: 237.1425).
sulting precipitates gave the crude product 8p (1.12g, 74%).
N,N″-[[[2-[(1,3-Benzodioxol-5-ylmethyl)amino]ethyl]-
Recrystallization from 2-PrOH gave analytically pure 8p as imino]di-2,1-ethanediyl]bis[N′-(tricyclo[3.3.1.13,7]dec-1-yl)-
colorless crystals.
thiourea] (13k) (Entry 19) Compound 13k was prepared
8p: mp 198–200°C (from 2-PrOH). IR (KBr) cm−1: 3275 from 1 (1.46g, 10.0mmol) with 6k (3.87g, 20.0mmol) and
(NH), 1548, 1542 (S=C–N), 1329 (CF3), 1111, 1067 (C=S). 2d (1.50g, 10.0mmol) under the conditions shown in Table
1H-NMR (DNSO-d6) δ: 2.80 (6H, t, J=6.7Hz, H2′), 3.57–3.90 S4. Separation of the products by flash chromatography
(6H, m, H1′), 7.62 (6H, d, J=8.4Hz, H3, 5), 7.70 (6H, d, (CH2Cl2 :95% EtOH:28% NH3=95:4.7:0.3→93:6.6:0.4) gave

Vol. 64, No. 12 (2016)
Chem. Pharm. Bull.
1777
13k (2.12g, 32%) as a white solid and 12k (0.903g, 15%) as a H2″), 3.56 (2H, s, Hα), 3.63 (2H, brs, H1″), 3.82 (6H, s,
pale yellow oil. CH3O on C3‴), 3.84 (6H, s, CH3O on C4‴), 5.87 (2H, s, H2),
13k: mp 48–50°C. IR (KBr) cm−1: 3276 (NH of sec-amine), 6.66–6.71 (2H, m, H7, 6), ca. 6.7 (1H, brs, NH), 6.73 (1H, brs,
1539 (C=S), 1247, 1038, 933 (C–O). 1H-NMR (CDCl3) δ: H4), 6.76 (2H, dd, J=8.2, 2.1Hz, H6‴), 6.82 (2H, d, J=8.2Hz,
1.67 (12H, brs, Ha), 2.09 (19H, brs, Hb, Hc, NH), 2.63–2.68 H5‴), 6.83 (2H, brs, H2‴), 7.88 (2H, brs, NH). 13C-NMR
(2H, m, H2′), 2.68–2.74 (6H, m, H1′, 2″), 3.56–3.64 (4H, m, (CDCl3) δ: 43.48 (C1″), 46.77 (C1′), 53.36 (Cα), 53.43 (C2′ or
H1″), 3.72 (2H, s, Hα), 5.93 (2H, s, H2), 6.08 (2H, brs, NH), 2″), 53.62 (C2″ or 2′), 56.09, 56.12 (OCH3), 100.89 (C2), 108.11
6.54 (2H, brs, NH), 6.73–6.79 (2H, m, H4, 6), 6.84 (1H, d, (C7 or 4), 108.59 (C4 or 7), 109.83 (C2‴), 111.65 (C5‴), 118.04
J=1.2Hz, H7). 13C-NMR (CDCl3) δ: 29.51 (Cc), 36.16 (Ca), (C6‴), 121.25 (C6), 129.69 (C1‴), 133.50 (C5), 146.61 (C3a
42.12 (Cb), 42.95 (C1″), 46.72 (C1′), 53.11 (C2″), 53.62 (Cα), or 7a), 147.72 (C4‴), 148.14 (C7a or 3a), 149.67 (C3‴), 181.27
53.80 (C2′), 54.07 (Cd), 100.88 (C2), 108.16 (C4), 108.73 (C=S). Positive-ion FAB-MS m/z: 671 (M+H+). HR-FAB-MS
(C7), 121.41 (C6), 133.61 (C5), 146.65 (C3a), 147.74 (C7a), m/z: 671.2684 (Calcd for C32H43N6O6S2: 671.2686). Anal. Calcd
180.77 (C=S). Positive-ion FAB-MS m/z: 667 (5, M+H+). HR- for C32H42N6O6S2·1.2H2O: C, 55.50; H, 6.46; N, 12.14. Found:
FAB-MS m/z: 667.3824 (Calcd for C36H55N6O2S2: 667.3828). C, 55.54; H, 6.24; N, 11.86.
(YO2-6) Anal. Calcd for C36H54N6O2S2·0.5H2O: C, 63.96; H,
8.20; N, 12.43. Found: C, 63.99; H, 8.19; N, 12.16.
N-[2-[Bis[2-[(1,3-benzodioxol-5-ylmethyl)amino]ethyl]-
amino]ethyl]-N′-[3-(trifluoromethyl)phenyl]thiourea (12o),
N-[2-[Bis[2-[(1,3-benzodioxol-5-ylmethyl)amino]ethyl]- N,N′-[[[2-[(1,3-Benzodioxol-5-ylmethyl)amino]ethyl]imino]-
amino]ethyl]-N′-(3,4-dimethoxyphenyl)thiourea (12l) and 1- di-2,1-ethanediyl]bis[N′-[3-(trifluoromethyl)phenyl]thio-
(3,4-Dimethoxyphenyl)-2-imidazolidinethione (16l) (Entry urea] (13o) and 1-[3-(Trifluoromethyl)phenyl]-2-imidazol-
20) Compound 12l was prepared from 1 (1.46g, 10.0mmol) idinethione (16o) (Entry 22) Compounds 12o and 13o were
with 6l (1.95g, 10.0mmol) and 2d (3.00g, 20.0mmol) under prepared from 1 (1.46g, 10.0mmol) with 6o (2.03g, 10.0mmol)
the conditions shown in Table S4. Separation of the prod- and 2d (3.00g, 20.0mmol) under the conditions shown in
ucts by flash chromatography (CH2Cl2 :95% EtOH:28% Table S4. Separation of the products by flash chromatography
NH3=93:6.6:0.4→85:14:1) gave 16l (trace) as a white solid, (CH2Cl2 :95% EtOH:28% NH3=93:6.6:0.4→85:14:1) gave
13l (1.50g, 22%) as a white solid, and 12l (2.77g, 45%) as a 16o (466mg, 19%) as a pale yellow semi-solid, 13o (1.51g,
white oil.
22%) as a pale yellow semi-solid, and 12o (3.42g, 55%) as a
12l: IR (KBr) cm−1: 3290 (NH of sec-amine), 1513 (C=S), pale yellow oil.
1242, 1034, 928 (C–O). 1H-NMR (CDCl3) δ: 2.76 (6H, t,
12o: IR (KBr) cm−1: 3260 (NH of sec-amine), 1490 (C=S),
1
J=5.8Hz, H2″, 2′), 2.82 (4H, t, J=5.8Hz, H1′), 3.75 (2H, t, 1332 (ArCF3), 1249, 1038, 930 (C–O). H-NMR (CDCl3) δ:
J=5.8Hz, H1″), 3.82 (3H, s, OCH3 on C4‴), 3.82 (2H, s, Hα), ca. 2.3 (4H, brs, NH), 2.58 (4H, t, J=5.5Hz, H2′), 2.65–2.70
3.83 (3H, s, OCH3 on C3‴), 4.48 (3H, brs, NH), 5.92 (4H, s, (6H, m, H1′, 2″), 3.60 (4H, s, Hα), 3.60 (2H, brs, H1″), 5.87
H2), 6.73 (2H, d, J=7.9Hz, H7), 6.76 (1H, d, J=8.5Hz, H5‴), (4H, s, H2), 6.67–6.71 (4H, m, H6, 7), 6.74 (2H, s, H4),
6.85 (2H, dd, J=7.9, 1.5Hz, H6), 6.91 (2H, d, J=1.5Hz, H4), 7.34–7.37 (2H, m, H4‴, 5‴), 7.56 (2H, brs, H2‴, 6‴). 13C-NMR
6.93 (1H, dd, J=8.5, 2.4Hz, H6‴), 7.26 (1H, brs, H2‴), 9.06 (CDCl3) δ: 43.24 (C1″), 46.66 (C1′), 52.57 (C2″), 53.46 (Cα),
(1H, brs, NH). 13C-NMR (CDCl3) δ: 42.12 (C1″), 45.25 (C1′), 53.95 (C2′), 100.95 (C2), 108.19 (C7), 108.57 (C4), 120.36
51.92 (Cα), 52.28 (C2′), 54.39 (C2″), 55.99, 56.08 (OCH3), (q, J=4.1Hz, C2‴), 121.30 (C6), 121.49 (q, J=4.1Hz, C4‴),
101.31 (C2), 108.55 (C7), 109.10 (C2‴), 109.60 (C4), 111.19 125.96 (q, J=272.1Hz, CF3), 127.05 (C6‴), 129.22 (C5‴),
(C5‴), 116.53 (C6‴), 123.14 (C6), 127.58 (C5), 132.14 (C1‴), 131.21 (q, J=33.1Hz, C3‴), 133.20 (C5), 139.29 (C1‴), 146.77
146.79 (C3‴), 147.93 (C7a), 148.14 (C3a), 148.90 (C4‴), 181.62 (C3a), 147.83 (C7a), 181.05 (C=S). Positive-ion FAB-MS
(C=S). Positive-ion FAB-MS m/z: 610 (M+H+). HR-FAB-MS m/z: 135 (100), 247 (14), 618 (6, M+H+). HR-FAB-MS m/z:
m/z: 610.2714 (Calcd for C31H40N5O6S: 610.2699). Anal. Calcd 618.2363 (Calcd for C30H35F3N5O4S: 618.2362). Anal. Calcd for
for C31H39N5O6S·0.9H2O: C, 59.48; H, 6.57; N, 11.19. Found: C30H34F3N5O4S·0.6H2O: C, 57.33; H, 5.65; N, 11.14. Found: C,
C, 59.51; H, 6.50; N, 11.09.
57.35; H, 5.65; N, 10.94.
16l: 13C-NMR (CDCl3) δ: 42.98 (C4), 53.24 (C5), 56.02,
13o: IR (KBr) cm−1: 3256 (NH of sec-amine), 1490 (C=S),
1
56.09 (OCH3), 109.47 (C2′), 111.70 (C5′), 117.53 (C6′), 130.34 1332 (ArCF3), 1251, 1038, 930 (C–O). H-NMR (CDCl3) δ:
(C1′), 147.69 (C4′), 149.41 (C3′), 181.30 (C=S). (Signals of 1.23 (3H, t, J=7.0Hz, CH3 of EtOH), ca. 2.15 (3H, brs, NH
only the predominant conformer were assigned.) Positive- and/or OH), 2.66–2.71 (2H, m, H2′), ca. 2.7 (1H, brs, NH or
ion FAB-MS m/z: 239 (M+H+). HR-FAB-MS m/z: 239.0850 OH), 2.71–2.78 (6H, m, H1′, 2″), 3.57 (2H, s, Hα), 3.69 (4H,
(Calcd for C11H15N2O2S: 239.0854).
brs, H1″), 3.70 (2H, q, J=7.0Hz, CH2 of EtOH), ca. 3.7 (1H,
N,N″-[[[2-[(1,3-Benzodioxol-5-ylmethyl)amino]ethyl]- brs, NH), 5.78 (2H, s, H2), 6.57–6.68 (3H, m, H6, 7, 4), ca.
imino]di-2,1-ethanediyl]bis[N′-(3,4-dimethoxyphenyl)thio- 6.65 (1H, brs, NH), 7.22–7.31 (4H, m, H4‴, 5‴), 7.36 (2H, brd,
urea] (13l) (Entry 21) Compound 13l was prepared from J=7.6Hz, H6‴), 7.59 (2H, s, H2‴). 13C-NMR (CDCl3) δ: 18.40
1 (731mg, 5.00mmol) with 6l (1.95g, 10.0mmol) and 2d (CH3 of EtOH), 43.36 (C1″), 46.82 (C1′), 53.29 (C2′), 53.59
(751mg, 5.00mmol) under the conditions shown in Table S4. (Cα), 53.66 (C2″), 58.34 (CH2 of EtOH), 100.99 (C2), 108.33
Separation of the products by flash chromatography (CH2Cl2: (C7), 108.61 (C4), 120.45 (q, J=4.1Hz, C2‴), 121.56 (C6),
95% EtOH:28% NH3=95:4.7:0.3→93:6.6:0.4) gave 8l 121.86 (q, J=4.1Hz, C4‴), 123.68 (q, J=272.1Hz, CF3), 127.07
(1.11g, 30%) as a white solid and 13l (1.15g, 34%) as a color- (C6‴), 129.25 (C5‴), 131.07 (q, J=30.0Hz, C3‴), 132.27 (C5),
less semisolid.
138.90 (C1‴), 146.93 (C3a), 147.85 (C7a), 181.20 (C=S). Posi-
13l: mp 65–69°C. IR (KBr) cm−1: 3264 (NH of sec-amine), tive-ion FAB-MS m/z: 135 (100), 247 (61), 687 (22, M+H+).
1
1512 (C=S), 1236, 1026, 927 (C–O). H-NMR (CDCl3) δ: 1.81 HR-FAB-MS m/z: 687.2006 (Calcd for C30H33F6N6O2S2:
(2H, brs, NH), 2.59 (4H, brs, H1′, 2′), 2.66 (4H, t, J=6.1Hz, 687.2011). Anal. Calcd for C30H32F6N6O2S2·EtOH: C, 52.45; H,

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Chem. Pharm. Bull.
Vol. 64, No. 12 (2016)
5.23; N, 11.47. Found: C, 52.44; H, 5.00; N, 11.46.
62.87; H, 7.33; N, 9.64.
N,N′-[[[2-[[Thioxo(3,4-dimethoxyphenylamino)methyl]-
16o: Positive-ion FAB-MS m/z: 135 (57), 247 (100, M+H+).
HR-FAB-MS m/z: 247.0515 (Calcd for C10H10F3N2S: 247.0517). amino]ethyl]imino]di-2,1-ethanediyl]bis[N-(1,3-benzodiox-
1H-NMR (CDCl3) δ: 2.70 (2H, t, J=4.8Hz, H5), 3.80 (2H, brd, ol-5-ylmethyl)acetamide] (14l) (Entry 25) Compound 14l
J=4.8Hz, H4), 7.20–7.24 (1H, m, H5′), 7.26 (1H, d, J=6.8Hz, was prepared from 12l (1.53g, 2.51mmol) and 4 (0.512g,
H4′), 7.50 (1H, brd, J=6.9Hz, H6′), 7.91 (1H, brs, NH), 7.97 5.02mmol) under the conditions shown in Table S5. Separation
(1H, brs, H2′). 13C-NMR (CDCl3) δ: 41.97 (C4), 52.57 (C5), of the product by centrifugal chromatography (CH2Cl2 :95%
119.65 (brs, C2′), 121.11 (C4′), 123.79 (q, J=273.1Hz, CF3), EtOH:28% NH3=95:4.7:0.3→93:6.6:0.4) gave 14l (1.39g,
126.27 (C6′), 129.11 (C5′), 130.66 (q, J=33.2Hz, C3′), 139.58 80%) as a white solid.
(C1′), 180.88 (C=S).
14l: mp 54–58°C. IR (KBr) cm−1: 1639 (C=O), 1512, 1490
1
General Procedure for the Preparation of CS-Symmetri- (C=S), 1237, 1036, 923 (C–O). H-NMR (CDCl3) δ: 2.03, 2.08,
cal Amide-Urea-Type TAEA Derivatives (14, 15): Example: 2.10*, 2.11 (6H, s, CH3CO), 2.45–2.6 (4H, m, H2′), 2.6–2.70
Synthesis of N-(1,3-Benzodioxol-5-ylmethyl)-N-[2-[Bis[2- (2H, m, H2″), ca. 2.65 (1H, brs, OH), 3.05–3.15, 3.24–3.3,
[[thioxo(tricyclo[3.3.1.13,7]dec-1-ylamino)methyl]amino]- 3.3–3.35* (4H, m, H1′), 3.57 (2H, brs, H1″), 3.80, 3.82*, 3.83
ethyl]amino]ethyl]acetamide (15k) (Entry 24) (Step 1) To (6H, s, OCH3), 4.33, 4.42*, 4.44 (4H, s, Hα), 5.91, 5.95*, 5.95
a pale yellow suspension of compound 13k (1.88g, 2.82mmol) (4H, s, H2), 6.55–6.95, 7.17–7.23 (9H, m, H6, 4, 7, 5‴, 6‴, 2‴),
in tetrahydrofuran (THF) (5mL) was added Ac2O (4, 288mg, 7.49, 8.30, 8.63* (2H, brs, NH). (The signals of the predomi-
2.82mmol) and the mixture was stirred for 30min at r.t. The nant conformer were asterisked.) 13C-NMR (CDCl3) δ: 21.64
crude precipitated solid was obtained as a white substance 15k (CH3CO), 42.46 (C1″), 43.61 (C1′), 51.53 (Cα), 51.88 (C2′),
(1.83g, 92%). Recrystallization from THF gave an analytically 52.74 (C2″), 55.71, 55.84 (OCH3), 100.99 (C2), 106.58 (C4),
pure sample as a white powder.
108.32 (C7), 108.91 (C2‴), 111.19 (C5‴), 116.22 (C6‴), 119.41
15k: mp 188–192°C (from THF). IR (KBr) cm−1: 1627 (C6), 129.92 (C5), 131.31 (C1‴), 146.96 (C3a or C7a), 147.72
(C=O), 1540 (C=S), 1357, 1238, 1038 (C–O). 1H-NMR (C4‴), 148.09 (C7a or C3a), 148.84 (C3‴), 171.32 (C=O),
(CDCl3) δ: 1.68 (12H, brs, Ha), 2.11 (6H, brs, Hc), 2.14 (12H, 181.02 (C=S). (Signals of only the predominant conformer
brs, Hb), 2.19 (3H, s, CH3), 2.63–2.68 (6H, m, H2′, 2″), 3.43 were assigned.) Positive-ion FAB-MS m/z: 694 (M+H+). HR-
(2H, t, J=5.5Hz, H1′), 3.55 (4H, q, J=5.2Hz, H1″), 4.53 FAB-MS m/z: 694.2928 (Calcd for C35H44N5O8S: 694.2911).
(2H, brs, Hα), 5.96 (2H, s, H2), 6.32 (2H, brs, NH), 6.62 Anal. Calcd for C35H43N5O8S·0.5H2O; C, 59.81; H, 6.31; N,
(2H, brs, NH), 6.63 (1H, d, J=7.9Hz, H6), 6.64 (1H, brs, 9.96. Found: C, 59.74; H, 6.31; N, 10.04.
H4), 6.7–6.8 (1H, d, J=7.9Hz, H7). 13C-NMR (CDCl3) δ:
N-(1,3-Benzodioxol-5-ylmethyl)-N-[2-[Bis[2-[[thioxo(3,4-
22.26 (CH3), 29.60 (Cc), 36.31 (Ca), 42.17 (Cb), 42.86 (C1″), dimethoxyphenylamino)methyl]amino] Ethyl]amino]ethyl]-
43.89 (C1′), 51.53 (Cα), 52.53 (C2′), 53.19 (C2″), 53.94 (Cd), acetamide (15l) (Entry 26) Compound 15l was prepared
101.27 (C2), 106.75 (C4), 108.65 (C7), 119.63 (C6), 129.72 from 13l (394mg, 587µmol) and 4 (59.9mg, 587µmol) under
(C5), 147.30 (C3a), 148.42 (C7a), 172.52 (C=O), 181.19 (C=S). the conditions shown in Table S5. Separation of the product
Positive-ion FAB-MS m/z: 709 (M+H+). HR-FAB-MS m/z: by centrifugal chromatography (CH2Cl2 :EtOH=95:5) gave
709.3948 (Calcd for C38H57N6O3S2: 709.3934). Anal. Calcd for 15l (368mg, 88%) as a white solid. An analytical sample of
C38H56N6O3S2·0.4H2O: C, 63.72; H, 7.99; N, 11.73. Found: C, 15l was obtained by recrystallization from dichloromethane
63.69; H, 8.13; N, 11.47.
(CH2Cl2) as an off-white solid.
N,N′-[[[2-[[Thioxo(tricyclo[3.3.1.13,7]dec-1-ylamino)meth-
15l: mp 193–194°C (from CH2Cl2). IR (KBr) cm−1: 1616
1
yl]amino]ethyl]imino]di-2,1-ethanediyl]bis[N-(1,3-benzodi- (C=O), 1511, 1453 (C=S), 1239, 1034, 922 (C–O). H-NMR
oxol-5-ylmethyl)acetamide] (14k) (Entry 23) Compound (DMSO-d6) δ: 1.99, 2.08* (3H, s, CH3CO), 2.54–2.66 (6H,
14k was prepared from 12k (1.22g, 2.00mmol) and Ac2O (4, m, H2′, 2″), 3.18–3.27 (2H, m, H1′), 3.49 (4H, brt, J=5.5Hz,
408mg, 4.00mmol) under the conditions shown in Table S5. H1″), 3.71 (6H, s, CH3 on C3‴), 3.72 (6H, s, CH3 on C4‴),
Separation of the product by flash chromatography (EtOAc : 4.39*, 4.41 (2H, brs, Hα), 5.70*, 6.00 (2H, s, H2), 6.64–6.98
EtOH=99:1) gave 14k (934mg, 68%) as a white solid.
(9H, m, H6, 7, 4, 6‴, 5‴, 2‴), 7.31 (2H, brs, NHβ), 9.39 (2H,
14k: mp 66–68°C. IR (KBr) cm−1: 3324 (NH), 1637 (C=O), s, NHγ). (The signals of the predominant conformer were as-
1
1490 (C=S), 1248, 1038, 924 (C–O). H-NMR (CDCl3) δ: 1.66 terisked.) 13C-NMR (DMSO-d6) δ: 21.15 (CH3CO), 41.94 (C1″),
(6H, brs, Ha), 2.06 (3H, brs, Hc), 2.12*, 2.16 (6H, s, CH3), 45.93 (C1′), 47.64 (Cα), 51.71 (C2′), 52.40 (C2″), 55.44, 55.68
2.08, 2.20* (6H, brs, Hb), 2.6–2.55 (4H, m, H2′), 2.6–2.7 (2H, (OCH3), 100.76 (C2), 107.93 (C7), 108.08 (C4), 109.30 (C2‴),
m, H2″), 3.2–3.25, 3.3–3.4* (4H, m, H1′), 3.4–3.5, 3.5–3.6* 111.98 (C5‴), 116.39 (C6‴), 120.96 (C6), 131.42 (C1‴ or C5),
(2H, m, H1″), 4.36, 4.45*, 4.48 (4H, s, Hα), 5.92, 5.96*, 5.97 132.12 (C5 or C1‴), 146.27 (C3a or C7a), 146.38 (C4‴), 147.29
(4H, s, H2), 6.34 (1H, brs, NH), 6.6–6.7 (4H, m, H4, 6), (C7a or C3a), 148.70 (C3‴), 169.64 (C=O), 180.26 (C=S).
6.7–6.8 (2H, m, H7), 7.18 (1H, brs, NH). (The signals of the (Signals of only the predominant conformer were assigned.)
predominant conformer were asterisked.) 13C-NMR (CDCl3) Positive-ion FAB-MS m/z: 713 (M+H+). HR-FAB-MS m/z:
δ: 21.84 (CH3), 29.51 (Cc), 36.30 (Ca), 41.88 (Cb), 42.41 (C1″), 713.2786 (Calcd for C34H45N6O7S2: 713.2791). Anal. Calcd for
43.38 (C1′), 51.34 (Cα), 51.99 (C2′), 52.69 (C2″), 53.27 (Cd), C34H44N6O7S2: C, 57.29; H, 6.22; N, 11.79. Found: C, 57.10; H,
101.09 (C2), 106.66 (C4), 108.50 (C7), 119.45 (C6), 130.07 6.17; N, 11.75.
(C5), 147.05 (C3a), 148.23 (C7a), 171.50 (C=O), 180.82 (C=S).
Preparation of C3-Symmetrical TAPA Derivatives (18a,
(Signals of only the predominant conformer were assigned.) 21) 2,2′,2″-[Nitrilotris(3,1-propanediyliminomethylene)]tris-
Positive-ion FAB-MS m/z: 692 (M+H+). HR-FAB-MS m/z: [6-methoxyphenol] (18a) Compound 18a was prepared
692.3488 (Calcd for C37H50N5O6S: 692.3482). Anal. Calcd for starting from the reaction of TAPA (17, 942mg, 5.0mmol)
C37H49N5O6S·0.9H2O: C, 62.76; H, 7.23; N, 9.89. Found: C, and 2a (3.04g, 20.0mmol) in a manner similar to that for

Vol. 64, No. 12 (2016)
Chem. Pharm. Bull.
1779
the preparation of compound 3a. The reactions in both cal assistance.
steps were conducted in EtOH instead of MeOH with the
ratio of TAPA:2a:NaBH4=1:4:8.2. Separation of the prod-
Conflict of Interest The authors declare no conflict of
uct by centrifugal chromatography (CHCl3 :2-PrOH:28% interest.
NH3=84:14:2) gave 18a (1.93g, 65%) as a light-brown semi-
solid.
Supplementary Materials The online version of this ar-
18a: mp 31–35°C. IR (KBr) cm−1: 3420 (OH), 3310 (NH), ticle contains supplementary materials.
1
1590, 1480 (C=C of Ar), 1235, 1075 (C–O). H-NMR (CDCl3) Table S1. Synthesis of C3-Symmetrical Amine-Type TAEA
δ: 1.63 (6H, qu, J=6.7Hz, H2′), 2.41 (6H, t, J=6.7Hz, H3′), Derivatives (3)
2.64 (6H, t, J=6.7Hz, H1′), 3.84 (9H, s, CH3O), 3.93 (6H, Table S2. Synthesis of C3-Symmetrical Amide-Type TAEA
s, Hα), 6.26 (6H, brs, NH, OH), 6.61 (3H, d, J=7.6Hz, H3), Derivatives (5)
6.71 (3H, dd, J=7.9, 7.6Hz, H4), 6.79 (3H, d, J=7.9Hz, H5). Table S3. Synthesis of C3-Symmetrical Urea-Type TAEA De-
13C-NMR (CDCl3) δ: 26.74 (C2′), 47.35 (C1′), 52.21 (C3′ or rivatives (7, 8)
Cα), 52.24 (Cα or C3′), 55.83 (CH3O), 110.83 (C5), 118.48 Table S4. Synthesis of CS-Symmetrical Amine-Urea-Type
(C4), 120.55 (C3), 122.69 (C2), 147.40 (C1), 147.96 (C6). TAEA Derivatives (12, 13)
Positive-ion FAB-MS m/z: 597 (M+H+). HR-FAB-MS m/z: Table S5. Synthesis of CS-Symmetrical Amide-Urea-Type
597.3644 (Calcd for C33H49N4O6: 597.3652). Anal. Calcd for TAEA Derivatives (14, 15)
C33H48N4O6·1.2 H2O: C, 64.10; H, 8.22; N, 9.06. Found: C,
64.15; H, 8.31; N, 8.91.
N, N′, N″-(Nitrilotri-3,1-propanediyl)tris[3,5-bis-
(trifluoromethyl)benzamide] (21) To
References and Notes
1) Humblet V., Misra P., Bhushan K. R., Nasr K., Ko Y.-S., Tsuka-
moto T., Pannier N., Frangioni J. V., Maison W., J. Med. Chem., 52,
544–550 (2009).
a solution of 17
(942mg, 5.0mmol) and triethylamine (TEA, 1.77g, 17.5mmol)
in CH2Cl2 (50mL) was added a solution of 3,5-bis(trifluoro-
methyl)benzoyl chloride (19, 4.78g, 17.3mmol) in CH2Cl2
(25mL) at 0°C under an N2 atmosphere. After stirring for
30min, the reaction mixture was stirred at r.t. for another
1h. The resulting mixture was poured into water (100mL),
and then the aqueous layer was extracted with CH2Cl2
(2×100mL). The combined organic layer was washed with
brine (50mL) and dried over MgSO4, and the solvent was
evaporated to give a yellow oil. Separation of the product
by flash chromatography (CH2Cl2 : 95% EtOH : 28% NH3=
97:2.7:0.3) and then recrystallization from MeOH gave com-
pound 21 (2.99g, 66%) as colorless crystals.
2) Contreras J.-M., Sippl W., “The Practice of Medicinal Chemistry,”
3rd ed., ed. by Wermuth C. G., Elsevier Academic Press, London,
2008, pp. 380–414 and related references cited therein.
3) Mibu N., Yokomizo K., Aki H., Ota N., Fujii H., Yuzuriha A.,
Saneyoshi S., Tanaka A., Koga A., Zhou J., Miyata T., Sumoto K.,
Chem. Pharm. Bull., 63, 935–944 (2015) and related references cited
therein.
4) Mibu N., Yokomizo K., Koga A., Honda M., Mizokami K., Fujii H.,
Ota N., Yuzuriha A., Ishimaru K., Zhou J., Miyata T., Sumoto K.,
Chem. Pharm. Bull., 62, 1032–1040 (2014) and related references
cited therein.
5) Furutachi M., Ejima A., Tsuru R., Goto S., Gondo T., Ako K., Fujii
S., Okumura A., Tozuka A., Yokomizo K., Zhou J., Inao H., Ono
Y., Kashige N., Miake F., Sumoto K., Heterocycles, 92, 925–935
(2016).
21: mp 205–206°C (from MeOH). IR (KBr) cm−1: 3265
1
(NH), 1645, 1550, 1280 (C=O), 1135 (CF3). H-NMR (DMSO-
6) Mibu N., Yokomizo K., Uchida W., Takemura S., Zhou J., Aki H.,
Miyata T., Sumoto K., Chem. Pharm. Bull., 60, 408–414 (2012).
7) Mibu N., Aki H., Ikeda H., Saito A., Uchida W., Yokomizo K.,
Zhou J., Miyata T., Sumoto K., J. Therm. Anal. Calorim., 113,
1015–1018 (2013).
d6) δ: 1.71(6H, t, J=7.0Hz, H2′), 2.49 (6H, m, H3′), 3.35 (6H,
m, H1′), 8.23 (3H, s, H4), 8.41 (6H, s, H2, H6), 8.94 (3H, t,
J=5.3Hz, NH). 13C-NMR (DMSO-d6) δ: 26.39 (C2′), 38.03
(C1′), 50.93 (C3′), 123.08 (CF3, q, J=273.1Hz), 124.58 (C4),
127.86 (C2, C6), 130.41 (C3, C5, q, J=33.1Hz), 136.68 (C1),
163.13 (C=0). Positive-ion FAB-MS m/z: 909 (M+H+). HR-
FAB-MS m/z: 909.2101 (Calcd for C36H31F18N4O3: 909.2109).
Anal. Calcd for C36H30F18N4O3: C, 47.59; H, 3.33; N, 6.17.
Found: C, 47.58; H, 3.34; N, 6.22.
Antiviral Activity Assay and Cytotoxicity of Synthesized
Trisubstituted Tris(aminoalkyl)amine Derivatives The an-
ti-HSV-1 activities (EC50) of the synthesized tris(aminoalkyl)-
amine derivatives were assessed by using a plaque reduction
assay17) and their cytotoxicity against Vero cells (CC50) was
also evaluated. The results are summarized in Tables 1, 2
together with data for aciclovir.18) Log P values for all of the
compounds for which biological activities were evaluated were
calculated by using CAChe v.6.1.12. There were few distinct
correlations between logP values and anti-HSV-1 activities
(EC50 values) among the compounds listed in Tables 1, 2.
8) In the field of peptide biomedicines, much attention is being paid
to interesting substances known as middle-size molecules (mid-size
molecules) having molecular weights of ca. 500–50000. New TAEA
derivatives of targeted symmetrical molecules have considerably
large molecular weights over 500; however, the mid-size TAEA
symmetrical derivatives that we have synthesized belong to a new
class of non-peptide compounds. Hitherto, we have already reported
that compound 20 has a lectin-like carbohydrate recognition prop-
erty and shows considerably high cytotoxic activity.6,7) Compound
20 and its analogues may be classified into the category of non-
peptide mid-size molecules (see also ref. 5). In terms of non-peptide
mid-size multivalent symmetrical molecules, a few new drugs such
as daclatasvir9) and ombitasvir10) that have recently been developed
for the treatment of hepatitis C virus (HCV) infection diseases can
be considered to be C2-symmetrical geometric non-peptide mid-size
molecules.
9) Belema M., Nguyen Van N., Bachand C., Deon D. H., Goodrich
J. T., James C. A., Lavoie R., Lopez O. D., Martel A., Romine J.
L., Ruediger E. H., Snyder L. B., St. Laurent D. R., Yang F., Zhu
J., Wong H. S., Langley D. R., Adams S. P., Cantor G. H., Chi-
malakonda A., Fura A., Johnson B. M., Knipe J. O., Parker D. D.,
Santone K. S., Fridell R. A., Lemm J. A., O’Boyle D. R. II, Colonno
R. J., Gao M., Meanwell N. A., Hamann L. G., J. Med. Chem., 57,
Acknowledgment The authors would like to thank Ms.
Ai Yuzuriha, Ms. Izumi Sakai, Ms. Marina Sano, Ms. Yuuna
Kawaguchi, Ms. Saki Sawai, Mr. Ryo Sato, Mr. Kenta Mori-
moto, and Mr. Shunsuke Shimomura for their valuable techni-

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Vol. 64, No. 12 (2016)
2013–2032 (2014).
trans–cis and trans–cis–cis). These expected structural properties
of full N-acetylation reaction products are considered to be the
most reasonable interpretation for the multiple splitting of carbon
signals assignable to the target molecules. For these complicated
13C-NMR spectroscopic data of N-acetylated TAEA derivatives, we
could not provide a successful assignment for the obtained product.
However, we detected the presence of four stereoisomers (ratio of
isomers=73:4:20:3) in the obtained N-acetylated compound 5c
by the HPLC method. The data obtained from IR (absorption band
10) DeGoey D. A., Randolph J. T., Liu D., Pratt J., Hutchins C., Donner
P., Krueger A. C., Matulenko M., Patel S., Motter C. E., Nelson L.,
Keddy R., Tufano M., Caspi D. D., Krishnan P., Mistry N., Koev
G., Reisch T. J., Mondal R., Pilot-Matias T., Gao Y., Beno D. W.
A., Maring C. J., Molla A., Dumas E., Campbell A., Williams L.,
Collins C., Wagner R., Kati W. M., J. Med. Chem., 57, 2047–2057
(2014).
11) Oshovsky G. V., Reinhoudt D. N., Verboom W., Angew. Chem. Int.
Ed., 46, 2366–2393 (2007).
1
of the amide group), HR-MS (correct molecular ions) and H-NMR
12) Tobey S. L., Anslyn E. V., J. Am. Chem. Soc., 125, 10963–10970
(2003).
13) Liu S., Yang L., Rettig J. S., Orvig C., Inorg. Chem., 32, 2773–2778
(1993).
measurements are fully consistent with the N-acetylated structures
shown in Tables S2 and S5. We used the obtained materials for fur-
ther biological experimental study without further purification.
15) Mann S., Nature (London), 365, 499–505 (1993).
14) In the 13C-NMR spectra of N-acetylated C3- and CS-type TAEA
derivatives (5a-e, 5b′, 14k, 14l, and 15l) except for 15k, multiple
carbon resonance signal patterns ascribable to characteristic car-
bons such as amide functionalities were observed. For example,
theoretically four cis–trans rotational isomers can be obtained from
16) Hosseini M. W., Coord. Chem. Rev., 240, 157–166 (2003).
17) Schinazi R. F., Peters J., Williams C., Chance D., Nahmias A. J.,
Antimicrob. Agents Chemother., 22, 499–507 (1982).
18) Ikeda T., Yokomizo K., Okawa M., Tsuchihashi R., Kinjo J., Nohara
T., Uyeda M., Biol. Pharm. Bull., 28, 1779–1781 (2005).
full N-acetylation of three secondary amines in the C3-type origi- 19) Calculated log P values by using CAChe v.6.1.12 for cyctotoxic
nal amine-type molecule 5c, i.e., two C3-symmetrical molecules
compounds 8p, 20 and 13o were 9.94, 8.30 and 7.40, respectively.
that have the same conformations (trans–trans–trans and cis–cis– 20) Compound 8k showed no significant anti-HSV-1 (EC50=>100 µM)
cis) with regard to three amide functionalities (=N–CO–CH3) in
the molecules and two CS-symmetrical N-acetylated compounds
having different combinations of conformational isomers (trans–
and cytotoxic (CC50=>200µM) activities, and calculated logP value
for this compound was 7.41. (see ref. 6).