Synthesis, antibacterial activities, and sustained perfume release properties of optically active5-hydroxy- And 5-acetoxyalkanethioamide analogues

Article abstract of DOI:10.1515/hc-2019-0103

5-Acetoxy- and 5-hydroxyalkanethioamide analogues showed high antibacterial activity against Staphylococcus aureus. Antibacterial thioamides were prepared from 5-alkyl-δ-lactones by amidation, thionation, and subsequent deacetylation. Optically active thioamides with 99% diastereomeric excesses were prepared by diastereomeric resolution using Cbz-L-proline as the resolving agent. Antibacterial thioamides were slowly lactonized by a lipase catalyst. Therefore, these thioamides are potential sustained-release perfume compounds having antibacterial properties.

Full text of DOI:10.1515/hc-2019-0103

Heterocycl. Commun. 2020; 26: 84–98
Research Article
Open Access
Yasutaka Shimotori*, Masayuki Hoshi, Narihito Ogawa, Tetsuo Miyakoshi and Taisei Kanamoto
Synthesis, antibacterial activities, and sustained
perfume release properties of optically active5-
hydroxy- and 5-acetoxyalkanethioamide
analogues
https://doi.org/10.1515/hc-2019-0103
However, the widespread use and abuse of antibiotics has
Received December 27, 2019; accepted April 14, 2020.
led to the emergence of various multidrug resistant strains
Abstract: 5-Acetoxy- and 5-hydroxyalkanethioamide and superbugs, resulting in a potential threat to human
analogues showed high antibacterial activity against health and survival. The rapid increase in multidrug
Staphylococcus aureus. Antibacterial thioamides were resistant pathogenic bacteria is surprising. Examples of
prepared from 5-alkyl-δ-lactones by amidation, thiona- multidrug-resistant bacteria include the methicillin-resis-
tion, and subsequent deacetylation. Optically active thio- tant Staphylococcus aureus (MRSA) and the vancomycin-
amides with 99% diastereomeric excesses were prepared resistant enterococci (VRE). S. aureus (SA) is a historically
by diastereomeric resolution using Cbz-L-proline as the major human pathogen that continues to be one of the
resolving agent. Antibacterial thioamides were slowly most common disease-causing bacteria in humans [3].
lactonized by a lipase catalyst. Therefore, these thioami- Every year, approximately 50% of the hospitalized pati-
des are potential sustained-release perfume compounds ents around the world suffer from nosocomial infections
having antibacterial properties.
due to drug-resistant bacteria [4]. Therefore, there is an
urgent requirement to develop new drugs and materi-
als that can combat drug-resistant bacteria [5-8]. Over
the past few decades, the treatment of bacterial infec-
tions has become more complicated due to their resis-
tance mechanism. In light of the increasing incidence of
antibiotic resistance, the research and development of
novel antimicrobial agents with excellent antibacterial
activity is highly desirable and is of great scientific sig-
nificance. We have synthesized various optically active
δ-lactones [9-11] and found that N-methyl-5-acetoxydo-
decanamide, N-methyl-5-acetoxytridecanamide, and
N-methyl-5-acetoxytetradecanamides, which are pre-
cursors of δ-lactones, have antibacterial activities [12].
N-Methyl-5-acetoxyalkanamides showed high antibac-
terial activities against Gram positive bacteria such as S.
aureus and Pneumococcus pneumoniae. It is known that
thioamide compounds have higher biological activities
than the corresponding amide compounds [13-18]. Fortuna
et al. and Xue et al. also reported that thioamide com-
pounds have higher antibacterial activity than the corres-
ponding amide compounds [19, 20]. Based on these facts,
we synthesized various N-methyl-5-acetoxyalkanethio-
amides and attempted to improve their antibacterial acti-
vities and establish the structure-activity relationship.
Keywords: thioamide, antibacterial activity, Staphylococ-
cus aureus, optically active, diastereomeric resolution
Introduction
In the past few decades, there has been growing concern
about the health hazards and infections caused by patho-
genic microorganisms [1]. Currently used antibiotics are
effective in treating numerous infectious diseases [2].
*Corresponding author: Yasutaka Shimotori, School of Regional
Innovation and Social Design Engineering, Kitami Institute of
Technology, 165 Koen-cho, Kitami, Hokkaido 090-8507, Japan,
Phone: +81(157)269307, Fax: +81(157)247719,
E-mail: yasu@mail.kitami-it.ac.jp
Masayuki Hoshi, School of Regional Innovation and Social Design
Engineering, Kitami Institute of Technology, 165 Koen-cho, Kitami,
Hokkaido 090-8507, Japan
Narihito Ogawa and Tetsuo Miyakoshi, Department of Applied
Chemistry, School of Science of Technology, Meiji University, 1-1-1
Higashi-mita, Tama-ku, Kawasaki 214-8571 Japan
Taisei Kanamoto, Faculty of Pharmaceutical Sciences, Showa
Pharmaceutical University, 3-3165 Higashi-Tamagawagakuen,
Machida 194-8543, Japan
Open Access. © 2020 Shimotori et al., published by De Gruyter.ꢀ
Attribution alone 4.0 License.
ꢀThis work is licensed under the Creative Commons

Y. Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties ꢁ
ꢁ85
In many cases, there is a difference in biological activity obtained in 40% yield. In order to improve the yield, thio-
between stereoisomers [21-25]. Therefore, optically active nation of 5-acetoxytetradecanamide was performed at 40,
N-methyl-5-acetoxyalkanethioamides were synthesized, 60, 80 and 100 °C for 5 min, respectively, and rac-2e was
and their antibacterial activities were evaluated. Further- obtained in 61%, 80%, 83%, and 62% yields. We decided to
more, N-methyl-5-acetoxyalkanethioamides are precur- perform the thionation of 5-acetoxyalkanamides at 80 °C
sors of δ-lactones. Many 5-alkyl-δ-lactones have a sweet for 5 min, as prolonged thionation at high temperature
and fruity odor [26-30]. Therefore, if fungal lipase cataly- reduced the yield. 5-Acetoxydodecanethioamide (rac-2c)
zes its lactonization, it can be used as a sustained-release and 5-acetoxytridecanethioamide (rac-2d) were obtai-
perfume compound having antibacterial properties. The ned in approximately 70% yields, and 5-acetoxyhexade-
lipase-catalyzed lactonization of alkanethioamide ana- canethioamide (rac-2f) was obtained in 99% yield. The
logues showing antibacterial activities was investigated.
deacetylation of N-methyl-5-acetoxyalkanethioamides
(rac-1) was then performed at 0 °C for 3 h using sodium
methoxide to give N-methyl-5-hydroxyalkanethioamides
(rac-3) in approximately 90% yield. However, deacetyla-
tion of 5-acetoxyalkanethioamides (rac-2) at 0 °C gave
5-hydroxyalkanethioamides (rac-4) in approximately 40%
yield. When the reaction was performed at -15 °C, rac-4
was obtained in approximately 70% yield, regardless of
the difference in the R1 group. The antibacterial activities
of the synthesized 5-acetoxyalkanethioamide analogues
(rac-1 and rac-2) and 5-hydroxyalkanethioamide ana-
logues (rac-3 and rac-4) were investigated (Table 1).
Results and discussion
Preparation of N-methyl-5-hydroxyalkanethioamides
(rac-3) and 5-hydroxyalkanethioamides (rac-4)
We had previously reported the synthesis of optically
active 5-alkyl-δ-lactones with >99% enantiomeric exces-
ses via diastereomeric resolution of racemic N-methyl-
5-hydroxyalkanamides using amino acid derivatives as the
optical resolving agent [9]. We hypothesized that optically
active 5-acetoxyalkanethioamides could be synthesized Antibacterial activities of racemic 5-aetoxy- and 5-hydro-
by the same method (Scheme 1). Therefore, diastereome-
ric resolution of 5-hydroxyalkanethioamides was planned
xyalkanethioamide analogues
using amino acid derivatives. N-Methyl-5-acetoxyalkana- The antibacterial activity value was calculated, com-
mides and 5-acetoxyalkanamides were prepared from the pounds with a value of 2.0 or more (≥99% killing ratio)
corresponding 5-alkyl-δ-lactones by a previous method were considered antibacterial substances (see experi-
[31]. Racemic 5-hydroxyalkanethioamides were prepared mental section). At antibacterial activity values of ≥7.0,
by thionation of 5-acetoxyalkanamides with Lawesson’s the bacteria were almost completely killed. Antibacte-
reagent and subsequent deacetylation under basic condi- rial activities against S. aureus and E. coli were tested,
tions. The thionation of N-methyl-5-acetoxyalkanamides and none of the thioamide compounds showed activities
was carried out using Lawesson’s reagent in toluene at against E. coli. Several thioamides showed high anti-
100 °C for 3 h. N-Methyl-5-acetoxyalkanethioamides (rac-1) bacterial activities against S. aureus (Table 1). Among
were obtained in approximately 90% yields, regardless of the N-methyl-5-acetoxyalkanethioamides (rac-1), of
the difference in the R1 group. In contrast, when 5-aceto- the primary thioamides rac-1b and rac-1c showed high
xytetradecanamide was thionized under the same con- activities, with rac-1c showing higher activity than rac-1b.
ditions, 5-acetoxytetradecanethioamide (rac-2e) was Among the 5-acetoxyalkanethioamides (rac-2), which are
Scheme 1 Synthesis of racemic 5-hydroxyalkanthioamide analogues.

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86ꢁ
Y.Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties
Table 1ꢂAntibacterial activities of racemic thioamide analogues against S. aureusa)
Antibacterial
Antibacterial
Sample
R1
R2
Sample
R1
R2
activity value [-]b)
activity value [-]b)
rac-1a
rac-1b
rac-1c
rac-1d
rac-1e
rac-1f
rac-2c
rac-2d
rac-2e
rac-2f
n-C5H11
n-C6H13
n-C7H15
n-C8H17
n-C9H19
n-C11H23
n-C7H15
n-C8H17
n-C9H19
n-C11H23
Me
<0.1
5.0
6.7
1.9
0.5
0.3
5.1
6.4
6.3
1.6
rac-3a
rac-3b
rac-3c
rac-3d
rac-3e
rac-3f
rac-4c
rac-4d
rac-4e
rac-4f
n-C5H11
n-C6H13
n-C7H15
n-C8H17
n-C9H19
n-C11H23
n-C7H15
n-C8H17
n-C9H19
n-C11H23
Me
<0.1
1.9
>7.0
6.4
0.5
0.3
5.1
4.3
4.8
<0.1
H
H
a) Final concentration of synthetic compounds is 500 μg/mL.
b) Data An antibacterial activity value of ≥2.0 indicates that the bacterial killing ratio is ≥99%.
secondary thioamides, all rac-2 except rac-2f showed high resolving agents [9, 32]. We found that Cbz-L-proline is an
antibacterial activity. The antibacterial activities appeared efficient resolving agent to obtain both the diastereomeric
toberelatedtothechainlength,as1band2c,and1cand2d, esters with high diastereomeric excesses. TLC analysis of
which have the same number of carbons respectively, the diastereomeric esters revealed a polarity difference,
showed similar activities. Moreover, high activities were regardless of the difference between R1 and R2 (5 and 6),
observed for rac-3c and rac-3d among N-methyl-5-hydro- and it was easy to obtain both the diastereomers with
xyalkanethioamides (rac-3). Among the tested thioamides over 99% diastereomeric excesses by silica gel column
in Table 1, rac-3c had the highest antibacterial activity. chromatography separation. We named the high polarity
These N-methyl-5-hydroxyalkanethioamides (rac-3c and diastereomers as 1st and the low polarity diastereomers
3d) showed higher activity than the corresponding as 2nd. Subsequently, each of the obtained diastereo-
N-methyl-5-acetoxyalkanethioamides (rac-1c and 1d). All meric esters (5 and 6) was deacylated to give optically
5-hydroxyalkanethioamides (rac-4) except rac-4f showed active 5-hydroxyalkanethioamide analogues (3 and 4)
antibacterial activities, which were lower than those of with approximately 90% and 70% yields for 3 and 4,
the corresponding 5-acetoxyalkanethioamides (rac-2). respectively. Acetylation of both 3 and 4 afforded opti-
From these results, it was found that an acetyl group is cally active 5-acetoxyalkanethioamide analogues (1 and 2)
not necessary for the expression of antibacterial activity in almost quantitatively. The absolute configuration at the
these thioamides. It is assumed that appropriate polarity C-5 position of the diastereomeric esters was determined
to penetrate the cell membrane of S. aureus is required to by 1H NMR, using the Trost method [33, 34]. Diastereomer
express antibacterial activities.
esters (7e and 8e) were prepared from optically active 3e
and 4e obtained by deacylation of 5e1st and 6e1st, using
(R)- or (S)-α-methoxyphenylacetic acid (Scheme 3). From
the 1H NMR analysis, the absolute configuration at the C-5
position in 5e1st and 6e1st was found to be the (R)-configu-
ration regardless of the difference in R2 group. Moreover,
Diastereomeric resolution of racemic 5-hydroxyalka-
nethioamide analogues (rac-3 and rac-4)
It is known that organic compounds having biological acti- no decrease in optical purity could be confirmed in the
vities differ in these activities between stereoisomers [21- deacylation of diastereomeric esters (5 and 6) under basic
25]. As thioamides showing antibacterial activities have an conditions. Antibacterial activity tests were carried out on
asymmetric center at the C-5 position, it is considered that these thioamide analogues (1-4).
there is a difference in antibacterial properties between
the stereoisomers. 5-Acetoxyalkanethioamide analogues
(1 and 2) and 5-hydroxyalkanethioamide analogues (3 Evaluation of antibacterial activities for optically active
5-acetoxy- and 5-hydroxyalkanethioamide analogues
and 4) with high antibacterial activities were converted to
the corresponding optically active forms (Scheme 2). We
have previously reported the diastereomeric resolution of Table 2 shows the results of antibacterial activity tests
various amide compounds using amino acid derivatives as against S. aureus for the optically active forms. There

Y. Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties ꢁ
ꢁ87
Scheme 2 Preparation of optically active thioamide analogues.
Scheme 3 Esterification of (R)-3e and 3e using (R)- or (S)-MPA.
were no great differences in the antibacterial activities Evaluation of lipase-catalyzed lactonization for use as a
sustained-release perfume
between the enantiomers of N-methyl-5-acetoxydode-
canethioamide (1c). In the 5-acetoxyalkanethioamides
(2), both 2d and 2e showed higher activities in (S)-form 5-Hydroxyalkanethioamide analogues which have anti-
than in (R)-form. On the other hand, in the N-methyl- bacterial activities against S. aureus are precursors of
5-hydroxyalkanethioamides (3) no great difference was 5-alkyl-δ-lactones. Many 5-alkyl-δ-lactones have a sweet
observed in the activity value between each enantiomer. and fruity odor [26-30]. We have also reported that
The antibacterial activity values of 5-hydroxyalkanethio- δ-tridecalactone has a walnut-like odor [11]. Therefore,
amides (4) tended to be slightly higher in the (R)-form as if a fungal-derived lipase catalyzes lactonization, it can
compared to the (S)-form. Although a difference in anti- be expected to be used as a sustained-release perfume
bacterial activity depending on the absolute configura- having antibacterial properties [35]. Lipase-catalyzed lac-
tion could be observed, the consistency of the absolute tonization was confirmed for 5-hydroxyalkanethioamide
configuration could not be confirmed. The antibacterial analogues (3c, 4d and 4e) that showed antibacterial acti-
activity values of (S)-2d and 3c at a final sample concen- vities (Scheme 4, Table 3). 5-Hydroxytridecanethioamide
tration of 500 μg/mL were equal to or greater than seven. (rac-4d) was stirred for 48 h at 40 °C in diethyl ether with
These samples were also tested for antibacterial activity each lipase derived from different sources. The yield of
at a final sample concentration of 200 μg/mL (noted in δ-tridecalactone(9d)was0.4%intheabsenceoflipase. On
brackets). The antibacterial activity of 3c decreased the other hand, all lipases catalyzed lactonization regard-
slightly in both enantiomers, but the activity of the (R)- less of the origin. All lipases showed no stereoselectivity
enantiomer was slightly higher than that of (S)-enantio- for lactonization. Although Novozym 435 gave the highest
mer. In contrast, 2d showed almost no reduction in the yield of 13.1%, the other lipases catalyzed lactonization
antibacterial activity values when the sample concen- with approximately 4-5% yields. All lipases similarly cata-
tration decreased. Of all the samples evaluated, (S)-2d lyzed lactonization for 5-hydroxytetradecanethioamide
showed the highest antibacterial activity.
(rac-4e) and N-methyl-5-hydroxydodecanethioamide

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88ꢁ
Y.Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties
Table 2ꢂAntibacterial activities of optically active thioamide analogues against S. aureusa)
Sample
Config.
Antibacterial
Sample
Config.
Antibacterial
activity value [-]b)
activity value [-]b)
1c
Racemate
5R
6.7
3d
Racemate
6.4 (6.4)c)
5.9 (5.9)c)
6.0 (6.1)c)
5.1
6.4
5R
5S
5S
7.0
2d
2e
3c
Racemate
5R
5S
Racemate
5R
5S
Racemate
6.4 (5.2)c)
5.8 (5.7)c)
>7.0 (>7.0)c)
6.3 (5.8)c)
6.3 (5.9)c)
6.3 (5.9)c)
>7.0 (6.4)c)
>7.0 (6.6)c)
>7.0 (6.3)c)
4c
4d
4e
Racemate
5R
5.9
5S
5.7
Racemate
5R
4.3
4.8
5S
4.2
Racemate
5R
4.8
5.3
5S
5.0
a) Final concentration of synthetic compounds is 500 μg/mL.
b) An antibacterial activity value of ≥2.0 indicates that the bacterial killing ratio is ≥99%.
c) Final concentration of synthetic compounds is 200 μg/mL.
(rac-3c), although with varying yields. The antibacte-
rial activity value was evaluated after incubation for 20
h. The yield after 48 h of lipase-catalyzed lactonization
was at most approximately 13%. It seems that if the cata-
lytic activity of lipase is strong, all thioamides convert to
lactones before they show antibacterial properties. Pre-
viously we have confirmed that δ-dodeca-, δ-trideca-,
and δ-tetradecalactone showed no antibacterial activities
against S. aureus. Catalytic lactonization would prefe-
rably be slow in order for these thioamides to exhibit anti-
bacterial activity. From these results, it was shown that
these thioamides showing antibacterial activities can be
expected to be used as a sustained-release perfume.
Scheme 4 Lipase-catalyzed lactonization of thioamide derivatives.
Experimental
General
NMR spectra were recorded on a JNM-ECA-600 spectrome-
ter (JEOL, Tokyo, Japan). Chemical shifts were reported in
parts per million with respect to internal TMS. Coupling
Conclusions
5-Acetoxy- and 5-hydroxyalkanethioamide analogues pre- constants (J) are quoted in Hz. Structural determination
pared from 5-alkyl-δ-lactones showed high antibacterial of all compounds was performed by the use of COSY,
activities against S. aureus. 5-Acetoxyalkanethioamide HMQC, and HMBC NMR techniques. Infrared spectra were
analogues (1 and 2) had a higher antibacterial activity in obtained on an FT-IR 460plus spectrometer (JASCO Corp.,
the (S)-form than in the (R)-form. Conversely, 5-hydroxyal- Tokyo, Japan). ESI high resolution mass spectra were pro-
kanethioamide analogues (3 and 4) showed only a slightly vided on an AccuTOF GCv 4G instrument (JEOL). Melting
higher antibacterial activity in the (S)-form than in the (R)- points (mp) were recorded on a MP-500D micro-melting-
form. Among the samples evaluated, (S)-2d showed the point apparatus from Yanaco Technical Science Co., Ltd.
highest antibacterial activity. In addition, lipase catalytic (Kyoto, Japan) and are uncorrected.
activity for these thioamides was tested. All lipases slowly
catalyzed the lactonization of thioamides regardless of
their origin. Therefore, we propose that these thioamides General procedure for thionation
can be used as sustained-release perfume compounds
having antibacterial activity.
To a solution of N-methyl-5-acetoxyalkanamides (1.0
mmol) or 5-acetoxyalkanethioamide (1.0 mmol) in dry

Y. Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties ꢁ
ꢁ89
Table 3ꢀEffect of source on lipase-catalyzed lactonizationa)
C-H), 2858 (CH2, C-H), 1734 (OC=O), 1541 (NHC=S), 1458
(C-H), 1375 (NHC=S), 1246 (C-C(=O)-O). 1H NMR (600 MHz,
CDCl3): δ 0.88 (t, J= 6.9 Hz, 3H, H-11), 1.26 (m, 8H, H-7, H-8,
H-9, H-10), 1.53 (m, 2H, H-6), 1.57 (m, 2H, H-4), 1.73 (m, 1H,
H-3), 1.84 (m, 1H, H-3), 2.05 (s, 3H, -OC(=O)CH3), 2.67 (m,
2H, H-2), 3.16 (d, J= 4.8 Hz, 3H, CH3NHC(=S)-), 4.85 (m, 1H,
H-5), 7.83 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): δ 14.1
(C-11), 21.4 (-OC(=O)CH3), 22.6 (-CH2-), 25.1 (C-3), 25.4 (-CH2-),
29.2 (-CH2-), 31.8 (-CH2-), 33.0 (CH3NHC(=S)-), 33.2 (C-4),
34.1 (C-6), 46.0 (C-2), 73.8 (C-5), 171.5 (-OC(=O)CH3), 205.8
(C-1). HRMS (FD). Calcd for C14H27NO2S (M): m/z 273.1762.
Found: m/z 273.1756.
N-Methyl-5-acetoxydodecanethioamide (1c) Yield:
94%; Colorless oil. Rf= 0.20 (eluent; n-hexane-EtOAc, 3:1,
v/v). IR (NaCl): cm-1 3254 (N-H), 2953 (CH3, C-H), 2928 (CH2,
C-H), 2856 (CH2, C-H), 1736 (OC=O), 1545 (NHC=S), 1458
(C-H), 1377 (NHC=S), 1244 (C-C(=O)-O). 1H NMR (600 MHz,
CDCl3): δ 0.88 (t, J= 6.9 Hz, 3H, H-12), 1.26 (m, 10H, H-7, H-8,
H-9, H-10, H-11), 1.53 (m, 2H, H-6), 1.56 (m, 2H, H-4), 1.73
(m, 1H, H-3), 1.84 (m, 1H, H-3), 2.05 (s, 3H, -OC(=O)CH3),
2.67 (m, 2H, H-2), 3.16 (d, J= 4.8 Hz, 3H, CH3NHC(=S)-), 4.85
(quin, J= 6.2 Hz, 1H, H-5), 7.80 (br s, 1H, NH). 13C NMR (150
MHz, CDCl3): δ 14.1 (C-12), 21.4 (-OC(=O)CH3), 22.7 (-CH2-),
25.1 (C-3), 25.4 (-CH2-), 29.2 (-CH2-), 29.5 (-CH2-), 31.8 (-CH2-),
33.0 (CH3NHC(=S)-), 33.2 (C-4), 34.1 (C-6), 46.0 (C-2), 73.8
(C-5), 171.5 (-OC(=O)CH3), 205.8 (C-1). HRMS (FD). Calcd for
Entry Substrate Lipase
Source
Yield [%]
1
2
3
rac-4d
None
-
0.4
13.1
3.8
Novozym 435 Candida antarctica B
Lipase AK
Pseudomonas
fluorescens
Aspergillus niger
4
Lipase AS
5.4
6.0
5.2
4.7
4.6
3.5
6.3
7.9
1.8
3.3
2.6
4.3
12.7
12.8
7.3
5
Lipase AYS
Candida cylindracea
6
Lipase F-AP15 Rhizopus oryzae
7
8
9
Lipase G
Lipase M
Lipase PS
PPL
Penicillium camemberti
Mucor javanicus
Burkholderia cepacia
Porcine pancreas
10
11
12
13
14
15
16
17
18
rac-4e
rac-3c
Novozym 435 Candida antarctica B
Lipase AS
Lipase G
Lipase PS
Aspergillus niger
Penicillium camemberti
Burkholderia cepacia
Novozym 435 Candida antarctica B
Lipase AS
Lipase G
Lipase PS
Aspergillus niger
Penicillium camemberti
Burkholderia cepacia
a) Substrate: 1.0 mmol, Novozym 435: 0.4 g, other lipase: 0.5 w/w,
Et2O: 20 mL, 40 °C, 48 h.
toluene (20 mL), Lawesson’s reagent (0.4 g, 1.0 mmol) was C15H29NO2S (M): m/z 287.1919. Found: m/z 287.1890.
added. The reaction mixture was heated for 3 h at 100 °C for N-Methyl-5-acetoxytridecanethioamide (1d) Yield:
the preparation of rac-1, and for 5 min at 80 °C for the pre- 90%; Colorless oil. Rf= 0.21 (eluent; n-hexane-EtOAc, 3:1,
paration of rac-2. After cooling, the solvent was evapora- v/v). IR (NaCl): cm-1 3252 (N-H), 2953 (CH3, C-H), 2927 (CH2,
ted under vacuum, and the remaining residue was applied C-H), 2856 (CH2, C-H), 1735 (OC=O), 1545 (NHC=S), 1458
to a chromatographic column on silica (eluent; n-hexane/ (C-H), 1377 (NHC=S), 1243 (C-C(=O)-O). 1H NMR (600 MHz,
EtOAc= 9/1) to afford the corresponding N-methyl-5-aceto- CDCl3): δ 0.88 (t, J= 7.6 Hz, 3H, H-13), 1.26 (m, 12H, H-7, H-8,
xyalkanethioamides (1) or 5-acetoxyalkanethioamides (2). H-9, H-10, H-11, H-12), 1.53 (m, 2H, H-6), 1.56 (m, 2H, H-4),
N-Methyl-5-acetoxydecanethioamide (1a) Yield: 1.73 (m, 1H, H-3), 1.84 (m, 1H, H-3), 2.05 (s, 3H, -OC(=O)CH3),
89%; Colorless oil. Rf= 0.19 (eluent; n-hexane-EtOAc, 3:1, 2.67 (m, 2H, H-2), 3.16 (d, J= 4.8 Hz, 3H, CH3NHC(=S)-), 4.85
v/v). IR (NaCl): cm-1 3255 (N-H), 2953 (CH3, C-H), 2929 (CH2, (quin, J= 6.2 Hz, 1H, H-5), 7.80 (br s, 1H, NH). 13C NMR (150
C-H), 2858 (CH2, C-H), 1732 (OC=O), 1543 (NHC=S), 1458 MHz, CDCl3): δ 14.1 (C-13), 21.4 (-OC(=O)CH3), 22.7 (-CH2-),
(C-H), 1375 (NHC=S), 1242 (C-C(=O)-O). 1H NMR (600 MHz, 25.1 (C-3), 25.4 (-CH2-), 29.3 (C-6), 29.5 (-CH2-), 31.9 (-CH2-),
CDCl3): δ 0.88 (t, J= 6.9 Hz, 3H, H-10), 1.28 (m, 6H, H-7, H-8, 33.0 (CH3NHC(=S)-), 33.2 (C-4), 34.1 (-CH2-), 46.0 (C-2), 73.8
H-9), 1.53 (m, 2H, H-6), 1.57 (m, 2H, H-4), 1.73 (m, 1H, H-3), (C-5), 171.5 (-OC(=O)CH3), 205.6 (C-1). HRMS (FD). Calcd for
1.84 (m, 1H, H-3), 2.06 (s, 3H, -OC(=O)CH3), 2.67 (m, 2H, C16H31NO2S (M): m/z 301.2075. Found: m/z 301.2075.
H-2), 3.16 (d, J= 4.8 Hz, 3H, CH3NHC(=S)-), 4.86 (m, 1H,
N-Methyl-5-acetoxytetradecanethioamide
(1e)
H-5), 7.91 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): δ 14.0 Yield: 89%; Colorless oil. Rf= 0.22 (eluent; n-hexane-
(C-10), 21.4 (-OC(=O)CH3), 22.6 (-CH2-), 25.1 (C-3), 31.7 (-CH2-), EtOAc, 3:1, v/v). IR (NaCl): cm-1 3254 (N-H), 2952 (CH3,
33.0 (CH3NHC(=S)-), 33.2 (C-4), 34.0 (C-6), 46.0 (C-2), 73.8 C-H), 2929 (CH2, C-H), 2856 (CH2, C-H), 1734 (OC=O), 1546
1
(C-5), 171.5 (-OC(=O)CH3), 205.8 (C-1). HRMS (FD). Calcd for (NHC=S), 1458 (C-H), 1377 (NHC=S), 1244 (C-C(=O)-O). H
C13H25NO2S (M): m/z 259.1606. Found: m/z 259.1608.
NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.9 Hz, 3H, H-14), 1.26
N-Methyl-5-acetoxyundecanethioamide (1b) Yield: (m, 14H, H-7, H-8, H-9, H-10, H-11, H-12, H-13), 1.53 (m, 2H,
91%; Colorless oil. Rf= 0.19 (eluent; n-hexane-EtOAc, 3:1, H-6), 1.56 (m, 2H, H-4), 1.73 (m, 1H, H-3), 1.84 (m, 1H, H-3),
v/v). IR (NaCl): cm-1 3256 (N-H), 2953 (CH3, C-H), 2930 (CH2, 2.05 (s, 3H, -OC(=O)CH3), 2.67 (m, 2H, H-2), 3.16 (d, J= 4.1

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Y.Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties
Hz, 3H, CH3NHC(=S)-), 4.85 (quin, J= 6.2 Hz, 1H, H-5), 7.84
(br s, 1H, NH). 13C NMR (150 MHz, CDCl3): δ 14.2 (C-14), 21.4
(-OC(=O)CH3), 22.7 (-CH2-), 25.1 (C-3), 25.4 (-CH2-), 29.3 (C-6),
29.5 (-CH2-), 29.6 (-CH2-), 31.9 (-CH2-), 33.0 (CH3NHC(=S)-),
33.2 (C-4), 34.1 (-CH2-), 46.0 (C-2), 73.8 (C-5), 171.5 (-OC(=O)
CH3), 205.8 (C-1). HRMS (FD). Calcd for C17H33NO2S (M): m/z
315.2232. Found: m/z 315.2191.
44.8 (C-2), 73.7 (C-5), 171.5 (-OC(=O)CH3), 210.4 (C-1). HRMS
(FD). Calcd for C15H29NO2S (M): m/z 287.1919. Found: m/z
2 8 7.1 8 7 7.
5-Acetoxytetradecanethioamide (2e) Yield: 83%;
Pale yellow oil. Rf= 0.19 (eluent; n-hexane-EtOAc, 3:1,
v/v). IR (NaCl): cm-1 3186 (N-H), 3309 (N-H), 2928 (CH3,
C-H), 2856 (CH2, C-H), 1716 (OC=O), 1456 (NHC=S), 1373
(NHC=S), 1258 (C-C(=O)-O). 1H NMR (600 MHz, DMSO-d6):
δ 0.86 (t, J= 6.9 Hz, 3H, H-14), 1.24 (m, 14H, H-7, H-8, H-9,
H-10, H-11, H-12, H-13), 1.49 (m, 4H, H-4, H-6), 1.63 (m, 2H,
H-3), 1.99 (s, 3H, -OC(=O)CH3), 2.45 (t, J= 7.3 Hz, 2H, H-2),
4.77 (quin, J= 5.5, 6.9 Hz, 1H, H-5), 9.05 (br s, 1H, NH), 9.24
(br s, 1H, NH). 13C NMR (150 MHz, DMSO-d6): δ 14.4 (C-14),
21.4 (-OC(=O)CH3), 22.6 (-CH2-), 24.9 (C-3), 25.2 (-CH2-), 29.2
(-CH2-), 29.3 (-CH2-), 29.4 (-CH2-), 31.8 (-CH2-), 33.2 (C-6),
34.0 (C-4), 44.5 (C-2), 73.6 (C-5), 170.6 (-OC(=O)CH3), 208.6
(C-1). HRMS (FD). Calcd for C16H31NO2S (M): m/z 301.2075.
Found: m/z 301.2033.
5-Acetoxyhexadecanethioamide (2f) Yield: 99%;
Colorless solid; Mp= 47-48 °C. Rf= 0.20 (eluent; n-hexane-
EtOAc, 3:1, v/v). IR (KBr): cm-1 3185 (N-H), 3310 (N-H), 2928
(CH3, C-H), 2855 (CH2, C-H), 1715 (OC=O), 1456 (NHC=S),
1374 (NHC=S), 1257 (C-C(=O)-O). 1H NMR (600 MHz,
DMSO-d6): δ 0.86 (t, J= 6.9 Hz, 3H, H-16), 1.24 (m, 18H, H-7,
N-Methyl-5-acetoxyhexadecanethioamide
(1f)
Yield: 90%; Pale yellow oil. Rf= 0.23 (eluent; n-hexane-
EtOAc, 3:1, v/v). IR (NaCl): cm-1 3252 (N-H), 2954 (CH3,
C-H), 2928 (CH2, C-H), 2856 (CH2, C-H), 1737 (OC=O), 1545
1
(NHC=S), 1458 (C-H), 1376 (NHC=S), 1244 (C-C(=O)-O). H
NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.9 Hz, 3H, H-16), 1.25
(m, 18H, H-7, H-8, H-9, H-10, H11, H-12, H-13, H-14, H-15),
1.53 (m, 2H- H-6), 1.56 (m, 2H, H-4), 1.72 (m, 1H, H-3), 1.84
(m, 2H, H-3), 2.05 (s, 3H, -OC(=O)CH3), 2.67 (m, 2H, H-2),
3.16 (d, J= 4.8 Hz, 3H, CH3NHC(=S)-), 4.85 (quin, J= 6.2 Hz,
1H, H-5), 7.80 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): δ
14.2 (C-16), 21.4 (-OC(=O)CH3), 22.7 (-CH2-), 25.1 (C-3), 25.5
(-CH2-), 29.4 (-CH2-), 29.5 (-CH2-), 29.6 (-CH2-), 29.6 (-CH2-),
29.7 (-CH2-), 29.7 (-CH2-), 32.0 (-CH2-), 33.0 (CH3NHC(=S)-),
33.2 (C-4), 34.1 (C-6), 46.0 (C-2), 73.8 (C-5), 171.5 (-OC(=O)
CH3), 205.9 (C-1). HRMS (FD). Calcd for C19H37NO2S (M): m/z
343.2545. Found: m/z 343.2479.
5-Acetoxydodecanethioamide (2c) Yield: 72%; Pale H-8, H-9, H-10, H-11, H-12, H-13, H-14, H-15), 1.48 (m, 4H,
yellow oil. Rf= 0.16 (eluent; n-hexane-EtOAc, 3:1, v/v). IR H-4, H-6), 1.62 (m, 2H, H-3), 1.99 (s, 3H, -OC(=O)CH3), 2.44
(NaCl): cm-1 3186 (N-H), 3310 (N-H), 2928 (CH3, C-H), 2856 (t, J= 7.3 Hz, 2H, H-2), 4.76 (quin, J= 6.0, 6.0 Hz, 1H, H-5),
(CH2, C-H), 1715 (OC=O), 1456 (NHC=S), 1375 (NHC=S), 9.11 (br s, 1H, NH), 9.32 (br s, 1H, NH). 13C NMR (150 MHz,
1
1259 (C-C(=O)-O). H NMR (600 MHz, CDCl3): δ 0.88 (t, J= DMSO-d6): δ 14.5 (C-16), 21.4 (-OC(=O)CH3), 22.6 (-CH2-),
6.9 Hz, 3H, H-12), 1.26 (m, 10H, H-7, H-8, H-9, H-10, H-11), 25.0 (C-3), 25.3 (-CH2-), 29.3 (-CH2-), 29.4 (-CH2-), 29.5 (-CH2-),
1.53 (m, 2H, H-6), 1.60 (m, 2H, H-4), 1.73 (m, 1H, H-3), 1.84 29.5 (-CH2-), 29.6 (-CH2-), 31.8 (-CH2-), 33.2 (C-6), 34.0 (C-4),
(m, 1H, H-3), 2.06 (s, 3H, -OC(=O)CH3), 2.64 (m, 1H, H-2), 2.70 44.5 (C-2), 73.6 (C-5), 170.6 (-OC(=O)CH3), 208.5 (C-1). HRMS
(m, 1H, H-2), 4.87 (quin, J= 6.9 Hz, 1H, H-5), 7.26 (br s, (FD). Calcd for C18H35NO2S (M): m/z 329.2389. Found: m/z
1H, NH), 7.80 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): δ 329.2340.
14.1 (C-12), 21.4 (-OC(=O)CH3), 22.7 (-CH2-), 25.0 (C-3), 25.5
(-CH2-), 29.2 (-CH2-), 29.5 (-CH2-), 31.8 (-CH2-), 33.2 (C-6), 34.2
(C-4), 44.7 (C-2), 73.8 (C-5), 171.5 (-OC(=O)CH3), 210.5 (C-1). General procedure for deacetylation of rac-1 and 2
HRMS (FD). Calcd for C14H27NO2S (M): m/z 273.1762. Found:
m/z 273.1734.
To a solution of rac-1 (1.0 mmol) or rac-2 (1.0 mmol)
5-Acetoxytridecanethioamide (2d) Yield: 73%; Pale in methanol (1.0 mL) was added a solution of sodium
yellow oil. Rf= 0.17 (eluent; n-hexane-EtOAc, 3:1, v/v). IR methoxide in methanol (5.0 M, 10 mL). The reaction
(NaCl): cm-1 3185 (N-H), 3309 (N-H), 2928 (CH3, C-H), 2856 mixture was stirred for 3 h at 0 °C for the deacetylation
(CH2, C-H), 1714 (OC=O), 1456 (NHC=S), 1375 (NHC=S), 1259 of rac-1, and for 30 min at -15 °C for the deacetylation of
1
(C-C(=O)-O). H NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.9 rac-2. The reaction was quenched by adding H2O (30 mL)
Hz, 3H, H-13), 1.26 (m, 12H, H-7, H-8, H-9, H-10, H-11, H-12), and CHCl3 (10 mL). The resultant mixture was extracted
1.53 (m, 2H, H-6), 1.60 (m, 2H, H-4), 1.73 (m, 1H, H-3), 1.84 with CHCl3, and the combined extracts were washed with
(m, 1H, H-3), 2.06 (s, 3H, -OC(=O)CH3), 2.64 (m, 1H, H-2), brine, dried over MgSO4, filtered and concentrated. Purifi-
2.69 (m, 1H, H-2), 4.87 (m, 1H, H-5), 7.29 (br s, 1H, NH), 7.87 cation of crude product by silica gel column chromatogra-
(br s, 1H, NH). 13C NMR (150 MHz, CDCl3): δ 14.1 (C-13), 21.4 phy (eluent; n-hexane/EtOAc= 1/1) gave the corresponding
(-OC(=O)CH3), 22.7 (-CH2-), 24.9 (-CH2-), 25.4 (-CH2-), 29.2 N-methyl-5-hydroxyalkanethioamides (rac-2) or 5-hydro-
(-CH2-), 29.5 (-CH2-×2), 31.8 (-CH2-), 33.1 (C-4), 34.1 (C-6), xyalkanethioamides (rac-4).

Y. Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties ꢁ
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N-Methyl-5-hydroxydecanethioamide (3a) Yield: (-CH2-), 33.1 (CH3NHC(=S)-), 35.9 (C-4), 37.8 (C-6), 46.3 (C-2),
92%; Colorless solid; Mp= 35-36 °C. Rf= 0.26 (eluent; 71.8 (C-5), 206.2 (C-1). HRMS (FD). Calcd for C14H29NOS (M):
n-hexane-EtOAc, 1:1, v/v). IR (KBr): cm-1 3243 (O-H, N-H), m/z 259.1970. Found: m/z 259.1972.
2928 (CH2, C-H), 2855 (CH2, C-H), 1539 (NHC=S), 1456 (C-H),
N-Methyl-5-hydroxytetradecanethioamide
(3e)
1
1378 (NHC=S). H NMR (600 MHz, CDCl3): δ 0.89 (t, J= 6.9 Yield: 90%; Colorless solid; Mp= 62-63 °C. Rf= 0.30 (eluent;
Hz, 3H, H-10), 1.30 (m, 5H, H-7, H-8, H-9), 1.44 (m, 4H, H-4, n-hexane-EtOAc, 1:1, v/v). IR (KBr): cm-1 3242 (O-H, N-H),
H-6, H-7), 1.52 (m, 1H, H-4), 1.89 (m, 2H, H-3), 2.31 (br s, 1H, 2929 (CH2, C-H), 2856 (CH2, C-H), 1538 (NHC=S), 1456 (C-H),
1
OH), 2.72 (m, 2H, H-2), 3.16 (d, J= 6.2 Hz, 3H, CH3NHC(=S)-), 1379 (NHC=S). H NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.9
3.63 (m, 1H, H-5), 8.12 (br s, 1H, NH). 13C NMR (150 MHz, Hz, 3H, H-14), 1.26 (m, 13H, H-7, H-8, H-9, H-10, H-11, H-12,
CDCl3): δ 14.1 (C-10), 22.7 (-CH2-), 25.4 (C-3), 25.5 (C-7), 31.9 H-13), 1.43 (m, 4H, H-4, H-6, H-7), 1.52 (m, 1H, H-4), 1.88 (m,
(-CH2-), 33.0 (CH3NHC(=S)-), 35.8 (C-4), 37.7 (C-6), 46.3 (C-2), 2H, H-3), 2.27 (br s, 1H, OH), 2.72 (m, 2H, H-2), 3.16 (d, J=
71.8 (C-5), 206.2 (C-1). HRMS (FD). Calcd for C11H23NOS (M): 4.8 Hz, 3H, CH3NHC(=O)-), 3.63 (m, 1H, H-5), 8.11 (br s, 1H,
m/z 217.1500. Found: m/z 217.1505.
NH). 13C NMR (150 MHz, CDCl3): δ 14.2 (C-14), 22.8 (-CH2-),
N-Methyl-5-hydroxyundecanethioamide
(3b) 25.6 (C-3), 29.4 (C-7), 29.7 (-CH2-), 29.8 (-CH2-), 29.9 (-CH2-),
Yield: 90%; Colorless solid; Mp= 40-41 °C. Rf= 0.27 32.0 (-CH2-), 33.1 (CH3NHC(=O)-), 35.9 (C-4), 37.8 (C-6), 46.4
(eluent; n-hexane-EtOAc, 1:1, v/v). IR (KBr): cm-1 3242 (O-H, (C-2), 71.9 (C-5), 206.2 (C-1). HRMS (FD). Calcd for C15H31NOS
N-H), 2928 (CH2, C-H), 2856 (CH2, C-H), 1539 (NHC=S), 1456 (M): m/z 273.2126. Found: m/z 273.2137.
(C-H), 1379 (NHC=S). 1H NMR (600 MHz, CDCl3): δ 0.88 (t,
N-Methyl-5-hydroxyhexadecanethioamide
(3f)
J= 6.9 Hz, 3H, H-11), 1.28 (m, 7H, H-7, H-8, H-9, H-10), 1.43 Yield: 91%; Colorless solid; Mp= 70-71 °C. Rf= 0.30 (eluent;
(m, 3H, H-4, H-6, H-7), 1.52 (m, 1H, H-4), 1.88 (m, 2H, H-3), n-hexane-EtOAc, 1:1, v/v). IR (KBr): cm-1 3242 (O-H, N-H),
2.51 (br s, 1H, OH), 2.72 (m, 2H, H-2), 3.16 (t, J= 4.8 Hz, 3H, 2928 (CH2, C-H), 2858 (CH2, C-H), 1539 (NHC=S), 1455 (C-H),
1
CH3NHC(=S)-), 3.62 (m, 1H, H-5), 8.27 (br s, 1H, NH). 13C 1379 (NHC=S). H NMR (600 MHz, CDCl3): δ 0.89 (t, J= 6.9
NMR (150 MHz, CDCl3): δ 14.2 (C-11), 22.7 (-CH2-), 25.6 (C-3), Hz, 3H, H-16), 1.26 (m, 17H, H-7, H-8, H-9, H-10, H-11, H-12,
25.8 (C-7), 29.4 (-CH2-), 31.9 (-CH2-), 33.0 (-CH2-), 35.9 (C-4), H-13, H-14, H-15), 1.44 (m, 4H, H-4, H-6, H-7), 1.53 (m, 1H,
37.7 (C-6), 46.3 (C-2), 71.8 (C-5), 206.1 (C-1). HRMS (FD). H-4), 1.89 (m, 2H, H-3), 2.22 (br s, 1H, OH), 2.72 (m, 2H, H-2),
Calcd for C12H25NOS (M): m/z 231.1657. Found: m/z 231.1651. 3.17 (d, J= 4.8 Hz, 3H, CH3NHC(=S)-), 3.63 (m, 1H, H-5), 8.07
N-Methyl-5-hydroxydodecanethioamide
(3c) (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): δ 14.2 (C-16), 22.8
Yield: 94%; Colorless solid; Mp= 53-54 °C. Rf= 0.27 (eluent; (-CH2-), 25.5 (C-3), 25.8 (C-7), 29.4 (-CH2-), 29.7 (-CH2-×3), 29.8
n-hexane-EtOAc, 1:1, v/v). IR (KBr): cm-1 3242 (O-H, N-H), (-CH2-×2), 32.0 (-CH2-), 33.0 (CH3NHC(=S)-), 35.8 (C-4), 37.8
2928 (CH2, C-H), 2855 (CH2, C-H), 1539 (NHC=S), 1373 (C-6), 46.3 (C-2), 71.9 (C-5), 206.2 (C-1). HRMS (FD). Calcd
1
(NHC=S). H NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.9 Hz, for C17H35NOS (M): m/z 301.2439. Found: m/z 301.2365.
3H, H-12), 1.28 (m, 9H, H-7, H-8, H-9, H-10, H-11), 1.43 (m,
5-Hydroxydodecanethioamide (4c) Yield: 71%;
3H, H-4, H-6, H-7), 1.52 (m, 1H, H-4), 1.89 (m, 2H, H-3), Colorless solid; Mp= 70-71 °C. Rf= 0.25 (eluent; n-hexane-
2.43 (br s, 1H, OH), 2.72 (m, 2H, H-2), 3.16 (d, J= 4.8 Hz, EtOAc, 1:1, v/v). IR (KBr): cm-1 3306 (N-H), 3152 (N-H), 2920
3H, CH3NHC(=S)-), 3.62 (m, 1H, H-5), 8.22 (br s, 1H, NH). (CH2, C-H), 2849 (CH2, C-H), 1456 (NHC=S), 1394 (NHC=S).
13C NMR (150 MHz, CDCl3): δ 14.2 (C-12), 22.7 (-CH2-), 25.6 1H NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.9 Hz, 3H, H-12),
(C-3), 25.8 (C-7), 29.4 (-CH2-), 29.7 (-CH2-), 31.9 (-CH2-), 33.1 1.28 (m, 9H, H-7, H-8, H-9, H-10, H-11), 1.41 (m, 1H, H-4),
(CH3NHC(=S)-), 35.9 (C-4), 37.8 (C-6), 46.3 (C-2), 71.8 (C-5), 1.44 (m, 3H, H-6, H-7), 1.55 (m, 1H, H-4), 1.88 (m, 2H, H-3),
206.2 (C-1). HRMS (FD). Calcd for C13H27NOS (M): m/z 2.49 (br s, 1H, OH), 2.71 (m, 2H, H-2), 3.64 (m, 1H, H-5), 7.72
245.1813. Found: m/z 245.1804.
(br s, 1H, NH), 8.07 (br s, 1H, NH). 13C NMR (150 MHz,
N-Methyl-5-hydroxytridecanethioamide
(3d) CDCl3): δ 14.2 (C-12), 22.7 (-CH2-), 25.4 (C-3), 25.8 (-CH2-), 29.4
Yield: 89%; Colorless solid; Mp= 54-55 °C. Rf= 0.27 (eluent; (-CH2-), 29.7 (-CH2-), 31.9 (-CH2-), 35.8 (C-6), 37.7 (C-4), 45.1
n-hexane-EtOAc, 1:1, v/v). IR (KBr): cm-1 3242 (O-H, N-H), (C-2), 71.8 (C-5), 210.8 (C-1). HRMS (FD). Calcd for C12H25NOS
2929 (CH2, C-H), 2856 (CH2, C-H), 1539 (NHC=S), 1456 (C-H), (M): m/z 231.1657. Found: m/z 231.1645.
1
1379 (NHC=S). H NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.9
5-Hydroxytridecanethioamide (4d) Yield: 73%;
Hz, 3H, H-13), 1.27 (m, 11H, H-7, H-8, H-9, H-10, H-11, H-12), Colorless solid; Mp= 72-73 °C. Rf= 0.26 (eluent; n-hexane-
1.43 (m, 3H, H-4, H-6, H-7), 1.52 (m, 1H, H-4), 1.89 (m, 2H, EtOAc, 1:1, v/v). IR (KBr): cm-1 3306 (N-H), 3153 (N-H), 2919
H-3), 2.41 (br s, 1H, OH), 2.72 (m, 2H, H-2), 3.16 (d, J= 4.8 Hz, (CH2, C-H), 2849 (CH2, C-H), 1456 (NHC=S), 1394 (NHC=S).
3H, CH3NHC(=S)-), 3.62 (m, 1H, H-5), 8.20 (br s, 1H, NH). 1H NMR (600 MHz, DMSO-d6): δ 0.88 (t, J= 6.8 Hz, 3H, H-13),
13C NMR (150 MHz, CDCl3): δ 14.2 (C-13), 22.8 (-CH2-), 25.6 1.27 (m, 16H, H-4, H-6, H-7, H-8, H-9, H-10, H-11, H-12), 1.65
(C-3), 25.8 (C-7), 29.4 (-CH2-), 29.7 (-CH2-), 29.8 (-CH2-), 32.0 (m, 1H, H-3), 1.77 (m, 1H, H-3), 2.47 (t, J= 6.9 Hz, 2H, H-2),

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Y.Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties
3.39 (m, 1H, H-5), 4.29 (d, J= 5.0 Hz, 1H, OH), 9.10 (br s,
(5R)-N-Methyl-5-[(2S)-N-(benzyloxy)carbonyl-
1H, NH), 9.29 (br s, 1H, NH). 13C NMR (150 MHz, DMSO- 2-pyrrolidinylmethanoyloxy]dodecanethioamide
d6): δ 14.5 (C-13), 22.7 (-CH2-), 25.7 (C-3), 25.8 (-CH2-), 29.3 [(5R,2’S)-5c1st] Yield: 47%; Colorless oil; [α]24D= -20.0
(-CH2-), 29.7 (-CH2-), 29.8 (-CH2-), 31.9 (-CH2-), 36.8 (C-6), 37.7 (c= 0.2, MeOH, 99% d.e.). Rf= 0.25 (eluent; n-hexane-
(C-4), 45.1 (C-2), 69.9 (C-5), 209.1 (C-1). HRMS (FD). Calcd for EtOAc, 3:1, v/v). IR (NaCl): cm-1 3308 (N-H), 2930 (CH2,
C13H27NOS (M): m/z 245.1813. Found: m/z 245.1805.
C-H), 2856 (CH2, C-H), 1742 (OC=O), 1659 (NC(=O)O), 1541
5-Hydroxytetradecanethioamide (4e) Yield: 69%; (NHC=S), 1456 (Ar, C=C), 1418 (NHC=S), 1120 (C-C(=O)-O),
1
Colorless solid; Mp= 76-77 °C. Rf= 0.27 (eluent; n-hexane- 698 (Ar, C-H). H NMR (600 MHz, CDCl3): (rotamer obser-
EtOAc, 1:1, v/v). IR (KBr): cm-1 3305 (N-H), 3152 (N-H), 2919 ved) δ 0.88 (t, J= 6.9 Hz, 3H, H-12), 1.20-1.32 (m, 10H, H-7,
(CH2, C-H), 2849 (CH2, C-H), 1456 (NHC=S), 1394 (NHC=S). H-8, H-9, H-10, H-11), 1.47-1.59 (m, 2H, H-6), 1.63-1.69 (m,
1H NMR (600 MHz, DMSO-d6): δ 0.88 (t, J= 6.6 Hz, 3H, 2H, H-4), 1.81-1.87 (m, 2H, H-3), 1.90-2.09 (m, 3H, -CH2- at
H-14), 1.27 (m, 18H, H-4, H-6, H-7, H-8, H-9, H-10, H-11, pyrrolidine), 2.23-2.32 (m, 1H, -CH2- at pyrrolidine), 2.46-
H-12, H-13), 1.66 (m, 1H, H-3), 1.76 (m, 1H, H-3), 2.47 (t, J= 2.52, 2.55-2.61 and 2.65-2.76 (m, 2H, H-2), 3.06 and 3.11 (d,
7.1 Hz, 2H, H-2), 3.39 (m, 1H, H-5), 4.20 (d, J= 5.0 Hz, 1H, J= 4.6 Hz, 3H, CH3NHC(=S)-), 3.47-3.57 and 3.62-3.69 (m, 2H,
OH), 9.04 (br s, 1H, NH), 9.22 (br s, 1H, NH). 13C NMR (150 -CH2- at pyrrolidine), 4.29-4.32 and 4.35-4.38 (dd, J= 4.0,
MHz, DMSO-d6): δ 14.4 (C-14), 22.6 (-CH2-), 25.7 (C-3), 25.8 4.0 Hz, 1H, -CH- at pyrrolidine), 4.82-4.87 and 4.99-5.04
(-CH2-), 29.2 (-CH2-), 29.5 (-CH2-), 29.7 (-CH2-), 29.8 (-CH2-), (m, 1H, H-5), 5.09 (d, J= 12.0 Hz, 1H, -C(=O)OCH2Ph), 5.18
31.8 (-CH2-), 36.9 (C-6), 37.7 (C-4), 45.1 (C-2), 69.9 (C-5), 209.2 (d, J= 12.0 Hz, 1H, -C(=O)OCH2Ph), 7.29-7.39 (m, 5H, Ph),
(C-1). HRMS (FD). Calcd for C14H29NOS (M): m/z 259.1970. 7.47 and 8.83 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3):
Found: m/z 259.1934.
(rotamer observed) δ 14.1 (C-12), 22.6 (-CH2-), 23.4 and 24.4
5-Hydroxyhexadecanethioamide (4f) Yield: 77%; (-CH2- at pyrrolidine), 24.7 and 25.4 (-CH2-), 25.5 (-CH2-),
Colorless solid; Mp= 85-86 °C. Rf= 0.29 (eluent; n-hexane- 29.2 (-CH2-), 29.3 (-CH2-), 30.0 (-CH2-), 31.1 and 31.7 (C-4),
EtOAc, 1:1, v/v). IR (KBr): cm-1 3306 (N-H), 3152 (N-H), 2920 32.9 and 33.1 (-CH2- at pyrrolidine), 33.9 and 35.1 (C-1’), 45.7
(CH2, C-H), 2848 (CH2, C-H), 1456 (NHC=S), 1393 (NHC=S). and 45.9 (C-2), 46.8 and 47.0 (-CH2- at pyrrolidine), 59.1 and
1H NMR (600 MHz, DMSO-d6): δ 0.86 (t, J= 7.3 Hz, 3H, H-16), 59.4 (-CH- at pyrrolidine), 66.9 and 67.2 (-C(=O)OCH2Ph),
1.24 (m, 22H, H-4, H-6, H-7, H-8, H-9, H-10, H-11, H-12, H-13, 74.1 and 74.6 (C-5), 127.4 and 127.7 (Ph), 128.0 and 128.2
H-14, H-15), 1.63 (m, 1H, H-3), 1.73 (m, 1H, H-3), 2.45 (t, (Ph), 128.5 and 128.6 (Ph), 136.1 and 136.5 (Ph), 154.3 and
J= 7.3 Hz, 2H, H-2), 3.36 (m, 1H, H-5), 4.18 (d, J= 5.5 Hz, 1H, 155.2 (-C(=O)OCH2Ph), 172.6 and 173.2 (-OC(=O)CH-), 205.7
OH), 9.02 (br s, 1H, NH), 9.20 (br s, 1H, NH). 13C NMR (150 (C-1). HRMS (FD). Calcd for C26H40N2O4S (M): m/z 476.2709.
MHz, DMSO-d6): δ 14.4 (C-16), 22.6 (-CH2-), 25.7 (C-3), 25.8 Found: m/z 476.2700.
(-CH2-), 29.2 (-CH2-), 29.5 (-CH2-), 29.6 (-CH2-×2), 29.7 (-CH2-),
(5S)-N-Methyl-5-[(2S)-N-(benzyloxy)carbonyl-
29.8 (-CH2-), 31.8 (-CH2-), 36.9 (C-6), 37.7 (C-4), 45.2 (C-2), 2-pyrrolidinylmethanoyloxy]dodecanethioamide
69.9 (C-5), 209.2 (C-1). HRMS (FD). Calcd for C16H33NOS (M): [(5S,2’S)-5c2nd] Yield: 49%; Colorless oil; [α]24D= -19.3
m/z 287.2283. Found: m/z 287.2238.
(c= 0.2, MeOH, 99% d.e.). Rf= 0.16 (eluent; n-hexane-
EtOAc, 3:1, v/v). IR (NaCl): cm-1 3304 (N-H), 2930
(CH2, C-H), 2856 (CH2, C-H), 1738 (OC=O), 1695 (NC(=O)O),
1541 (NHC=S), 1456 (Ar, C=C), 1418 (NHC=S), 1120 (C-C(=O)-
General procedure for esterification of rac-3 and 4
1
O), 696 (Ar, C-H). H NMR (600 MHz, CDCl3): (rotamer
To a solution of rac-3 or 4 (1.0 mmol) in THF (10 mL) observed) δ 0.88 (t, J= 6.8 Hz, 3H, H-12), 1.21-1.32 (m, 10H,
were added 1-ethyl-3-(3-dimethylaminopropyl)carbodii- H-7, H-8, H-9, H-10, H-11), 1.41-1.78 (m, 6H, H-3, H-4, H-6),
mide (EDCI) (0.31 g, 2.0 mmol), 4-dimethylaminopyridine 1.89-1.99 (m, 2H, -CH2- at pyrrolidine), 2.01-2.08 (m, 1H,
(DMAP) (0.12 g, 1.0 mmol), and N-carbobenzoxy-L-proline -CH2- at pyrrolidine), 2.22-2.32 (m, 1H, -CH2- at pyrrolidine),
(0.50 g, 2.0 mmol) at 0 °C. The reaction mixture was stirred 2.47-2.58 and 2.65-2.71 (m, 2H, H-2), 3.13 (t, J= 4.6 Hz, 3H,
for 2 h at 0 °C, and then for 3 h at room temperature. The CH3NHC(=S)-), 3.50-3.57 (m, 1H, -CH2- at pyrrolidine), 3.58-
reaction was quenched with aqueous 0.1 M HCl (5 mL). The 3.64 (m, 1H, -CH2- at pyrrolidine), 4.37-4.40 (m, 1H, -CH- at
resultant mixture was extracted with EtOAc, and the com- pyrrolidine), 4.80-4.85 and 4.91-4.96 (m, 1H, H-5), 5.05-5.18
bined extracts were washed with brine, dried over MgSO4, (m, 2H, -C(=O)OCH2Ph), 7.28-7.38 (m, 5H, Ph), 7.68 and 8.31
filtered, and concentrated. Flash column chromatography (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): (rotamer obser-
(eluent; n-hexane/EtOAc= 3/1) gave the corresponding 51st ved) δ 14.1 (C-12), 22.6 (-CH2-), 23.5 and 24.1 (C-4), 24.4 and
and 52nd or 61st and 62nd, respectively.
24.5 (-CH2-), 25.3 and 25.4 (-CH2-), 29.1 (-CH2-), 29.4 (-CH2-),

Y. Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties ꢁ
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30.3 and 31.0 (-CH2-), 31.7 and 31.8 (-CH2-), 32.7 (C-4), 33.0 -CH2- at pyrrolidine), 4.36-4.41 (m, 1H, -CH- at pyrrolidine),
and 33.1 (-CH2- at pyrrolidine), 33.7 and 34.5 (CH3NHC(=S)-), 4.79-4.85 and 4.90-4.96 (m, 1H, H-5), 5.05-5.18 (m, 2H,
45.9 (C-2), 46.7 and 47.0 (-CH2- at pyrrolidine), 59.4 and 60.0 -OCH2Ph), 7.28-7.39 (m, 5H, Ph), 7.71 and 8.31 (br s, 1H, NH).
(-CH- at pyrrolidine), 66.9 and 67.0 (-OCH2Ph), 74.2 and 74.7 13C NMR (150 MHz, CDCl3): (rotamer observed) δ 14.1 (C-13),
(C-5), 127.4 and 127.7 (Ph), 128.1 and 128.2 (Ph), 128.5 and 22.7 (-CH2-), 23.5 and 24.2 (-CH2- at pyrrolidine), 24.5 (-CH2-),
128.6 (Ph), 136.3 and 136.4 (Ph), 154.6 and 155.1 (-NC(=O) 25.3 (-CH2-), 29.2 (-CH2-), 29.4 (-CH2-), 30.3 (-CH2-), 31.0
OCH2Ph), 172.5 and 172.7 (-OC(=O)CH-), 205.4 (C-1). HRMS and 31.8 (-CH2-), 32.7 (C-4), 33.1 and 32.9 (-CH2- at pyrroli-
(FD). Calcd for C26H40N2O4S (M): m/z 476.2709. Found: m/z dine), 33.7 and 34.5 (CH3NHC(=S)-), 45.9 (C-2), 46.7 and 47.0
476.2697.
(-CH2- at pyrrolidine), 59.4 and 60.0 (-CH- at pyrrolidine),
(5R)-N-Methyl-5-[(2S)-N-(benzyloxy)carbonyl- 67.0 (-OCH2Ph), 74.3 and 74.7 (C-5), 127.4 and 127.7 (Ph),
2-pyrrolidinylmethanoyloxy]tridecanethioamide 128.0 and 128.2 (Ph), 128.5 and 128.6 (Ph), 136.3 and 136.4
[(5R,2’S)-5d1st] Yield: 46%; Colorless oil; [α]20D= -36.2 (c= (Ph), 154.6 and 155.1 (-NC(=O)OCH2Ph), 172.5 and 172.7
0.2, MeOH, 99% d.e.). Rf= 0.24 (eluent; n-hexane-EtOAc, (-OC(=O)CH-), 205.5 and 205.6 (C-1). HRMS (FD). Calcd for
3:1, v/v). IR (NaCl): cm-1 3308 (N-H), 2929 (CH2, C-H), 2856 C27H42N2O4S (M): m/z 490.2865. Found: m/z 490.2803.
(CH2, C-H), 1742 (OC=O), 1658 (NC(=O)O), 1542 (NHC=S),
(5R)-5-[(2S)-N-(benzyloxy)carbonyl-2-pyrrolidi-
1456 (Ar, C=C), 1418 (NHC=S), 1119 (C-C(=O)-O), 698 (Ar, nylmethanoyloxy]dodecanethioamide [(5R,2’S)-6c1st]
1
C-H). H NMR (600 MHz, CDCl3): (rotamer observed) δ Yield: 40%; Colorless oil; [α]18D= -39.1 (c= 0.2, MeOH, 99%
0.88 (t, J= 7.3 Hz, 3H, H-13), 1.25-1.31 (m, 12H, H-7, H-8, H-9, d.e.). Rf= 0.15 (eluent; n-hexane-EtOAc, 3:1, v/v). IR (NaCl):
H-10, H-11, H-12), 1.41-1.62 (m, 2H, H-6), 1.64-1.71 (m, 2H, cm-1 3312 (N-H), 3204 (N-H), 2930 (CH2, C-H), 2856 (CH2,
H-4), 1.80-1.88 (m, 2H, H-3), 1.92-2.08 (m, 3H, -CH2- at pyr- C-H), 1740 (OC=O), 1693 (NC(=O)O), 1418 (NHC=S), 1120
1
rolidine), 2.22-2.33 (m, 1H, -CH2- at pyrrolidine), 2.46-2.60 (C-C(=O)-O), 696 (Ar, C=C). H NMR (600 MHz, CDCl3):
and 2.64-2.76 (m, 2H, H-2), 3.06 and 3.10 (d, J= 4.6 Hz, 3H, (rotamer observed) δ 0.88 (t, J= 6.9 Hz, 3H, H-12), 1.23-1.31
CH3NHC(=S)-), 3.50-3.58 (m, 1H, -CH2- at pyrrolidine), 3.61- (m, 10H, H-7, H-8, H-9, H-10, H-11), 1.42-1.59 (m, 2H, H-6),
3.71 (m, 1H, -CH2- at pyrrolidine), 4.30 and 4.37 (dd, J= 4.6, 1.64-1.68 (m, 2H, H-4), 1.80-1.87 (m, 2H, H-3), 1.90-1.99 (m,
4.1 Hz, 1H, -CH- at pyrrolidine), 4.81-4.87 and 4.98-5.04 (m, 2H, -CH2- at pyrrolidine), 2.01-2.08 (m, 1H, -CH2- at pyrroli-
1H, H-5), 5.09 (d, J= 12.4 Hz, 1H, -OCH2Ph), 5.18 (d, J= 12.4 dine), 2.23-2.29 (m, 1H, -CH2- at pyrrolidine), 2.45-2.49 and
Hz, 1H, -OCH2Ph), 7.29-7.40 (m, 5H, Ph), 7.52 and 8.81 (br 2.61-2.66 (m, 1H, H-2), 2.53-2.58 and 2.72-2.77 (m, 1H, H-2),
s, 1H, NH). 13C NMR (150 MHz, CDCl3): (rotamer observed) 3.50-3.56 (m, 1H, -CH2- at pyrrolidine), 3.61-3.67 (m, 1H,
δ 14.2 (C-13), 22.7 (-CH2-), 24.5 (-CH2- at pyrrolidine), 25.4 -CH2- at pyrrolidine), 4.29 and 4.37 (dd, J= 8.6, 4.1 Hz, 1H,
(-CH2-), 25.5 (-CH2-), 29.3 (-CH2-), 29.4 (-CH2-), 29.5 (-CH2-), -CH- at pyrrolidine), 4.85 and 4.99 (quin, J= 5.5, 6.9 Hz, 1H,
30.1 (-CH2-), 31.9 (C-4), 33.2 (-CH2- at pyrrolidine), 35.2 H-1), 5.05 and 5.16 (d, J= 12.4 Hz, 1H, -C(=O)OCH2Ph), 5.10
(CH3NHC(=S)-), 45.9 (C-2), 46.9 (-CH2- at pyrrolidine), 59.5 and 5.20 (d, J= 12.4 Hz, 1H, -C(=O)OCH2Ph), 7.31-7.33 (m, 2H,
(-CH- at pyrrolidine), 67.2 (-OCH2Ph), 74.2 (C-5), 127.5 and Ph), 7.36-7.38 (m, 3H, Ph), 7.09 and 7.41 (br s, 1H, NH), 7.51
127.7 (Ph), 128.3 (Ph), 128.5 and 128.7 (Ph), 136.2 (Ph), 155.2 and 8.34 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): (rotamer
(-NC(=O)OCH2Ph), 172.7 (-OC(=O)CH-), 205.8 (C-1). HRMS observed) δ 14.2 (C-12), 22.7 (-CH2-), 24.5 and 24.7 (-CH2- at
(FD). Calcd for C27H42N2O4S (M): m/z 490.2865. Found: m/z pyrrolidine), 25.0 (-CH2-), 25.3 and 25.4 (-CH2-), 29.3 (-CH2-),
490.2816.
29.4 (-CH2-), 30.1 (-CH2- at pyrrolidine), 31.1 and 31.8 (-CH2-),
(5S)-N-Methyl-5-[(2S)-N-(benzyloxy)carbonyl- 34.0 and 35.0 (-CH2-), 44.4 and 44.8 (C-2), 46.8 and 47.1
2-pyrrolidinylmethanoyloxy]tridecanethioamide (-CH2- at pyrrolidine), 59.2 and 59.5 (-CH- at pyrrolidine),
[(5S,2’S)-5d2nd] Yield: 46%; Colorless oil; [α]20D= -41.7 (c= 67.1 and 67.3 (-C(=O)OCH2Ph), 74.1 and 74.7 (C-5), 127.4 and
0.2, MeOH, 99% d.e.). Rf= 0.15 (eluent; n-hexane-EtOAc, 127.8 (Ph), 128.0 and 128.2 (Ph), 128.6 and 128.7 (Ph), 136.4
3:1, v/v). IR (NaCl): cm-1 3303 (N-H), 2929 (CH2, C-H), 2856 (Ph), 155.2 (-NC(=O)OCH2Ph), 172.9 (-OC(=O)CH-), 210.9
(CH2, C-H), 1738 (OC=O), 1695 (NC(=O)O), 1541 (NHC=S), (C-1). HRMS (FD). Calcd for C25H38N2O4S (M): m/z 462.2552.
1455 (Ar, C=C), 1418 (NHC=S), 1120 (C-C(=O)-O), 696 (Ar, Found: m/z 462.2534.
C-H). 1H NMR (600 MHz, CDCl3): (rotamer observed) δ 0.88
(5S)-5-[(2S)-N-(Benzyloxy)carbonyl-2-pyrrolidi-
(t, J= 6.9 Hz, 3H, H-13), 1.19-1.33 (m, 12H, H-7, H-8, H-9, H-10, nylmethanoyloxy]dodecanethioamide [(5S,2’S)-6c2nd
]
H-11, H-12), 1.40-1.80 (m, 6H, H-3, H-4, H-6), 1.88-1.99 (m, Yield: 40%; Colorless oil; [α]19D= -40.5 (c= 0.2, MeOH,
2H, -CH2- at pyrrolidine), 2.00-2.08 (m, 1H, -CH2- at pyrro- 99% d.e.). Rf= 0.08 (eluent; n-hexane-EtOAc, 3:1, v/v). IR
lidine), 2.21-2.31 (m, 1H, -CH2- at pyrrolidine), 2.46-2.57 and (NaCl): cm-1 3310 (N-H), 3204 (N-H), 2928 (CH2, C-H), 2856
2.64-2.71 (m, 2H, H-2), 3.13 (t, J= 4.6 Hz, 3H, CH3NHC(=S)-), (CH2, C-H), 1740 (OC=O), 1693 (NC(=O)O), 1416 (NHC=S),
3.50-3.56 (m, 1H, -CH2- at pyrrolidine), 3.57-3.64 (m, 1H, 1120 (C-C(=O)-O), 696 (Ar, C=C). 1H NMR (600 MHz, CDCl3):

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Y.Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties
(rotamer observed) δ 0.87 (t, J= 6.9 Hz, 3H, H-12), 1.21-1.32
(5S)-5-[(2S)-N-(Benzyloxy)carbonyl-2-pyrrolidi-
(m, 10H, H-7, H-8, H-9, H-10, H-11), 1.45-1.77 (m, 6H, H-3, nylmethanoyloxy]tridecanethioamide [(5S,2’S)-6d2nd
]
H-4, H-6), 1.88-2.00 (m, 2H, -CH2- at pyrrolidine), 2.02-2.07 Yield: 47%; Colorless oil; [α]20D= -34.9 (c= 0.2, MeOH,
(m, 1H, -CH2- at pyrrolidine), 2.22-2.31 (m, 1H, -CH2- at pyr- 99% d.e.). Rf= 0.09 (eluent; n-hexane-EtOAc, 3:1, v/v). IR
rolidine), 2.46-2.51 (m, 1H, H-2), 2.53-2.58 and 2.64-2.68 (m, (NaCl): cm-1 3310 (N-H), 3202 (N-H), 2928 (CH2, C-H), 2855
1H, H-2), 3.50-3.55 (m, 1H, -CH2- at pyrrolidine), 3.59-3.62 (CH2, C-H), 1740 (OC=O), 1693 (NC(=O)O), 1417 (NHC=S),
(m, 1H, -CH2- at pyrrolidine), 4.36-4.39 (m, 1H, -CH- at pyr- 1120 (C-C(=O)-O), 696 (Ar, C=C). 1H NMR (600 MHz, CDCl3):
rolidine), 4.82-4.86 and 4.92-4.96 (m, 1H, H-5), 5.12 (m, 2H, (rotamer observed) δ 0.88 (t, J= 7.6 Hz, 3H, H-13), 1.21-1.29
-C(=O)OCH2Ph), 7.29-7.38 (m, 5H, Ph), 7.54 (br s, 1H, NH), (m, 12H, H-7, H-8, H-9, H-10, H-11, H-12), 1.44-1.77 (m, 6H,
7.92 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): (rotamer H-3, H-4, H-6), 1.88-2.00 (m, 2H, -CH2- at pyrrolidine), 2.02-
observed) δ 14.2 (C-12), 22.7 (-CH2-), 23.6 and 24.2 (-CH2-), 2.07 (m, 1H, -CH2- at pyrrolidine), 2.22-2.31 (m, 1H, -CH2- at
24.4 and 24.5 (-CH2- at pyrrolidine), 25.3 and 25.4 (-CH2-), pyrrolidine), 2.46-2.51 (m, 1H, H-2), 2.53-2.58 and 2.63-2.68
29.2 (-CH2-), 29.4 (-CH2-), 30.3 and 31.1 (-CH2- at pyrrolidine), (m, 1H, H-2), 3.50-3.55 (m, 1H, -CH2- at pyrrolidine), 3.59-
31.8 (-CH2-), 32.8 and 32.9 (-CH2-), 33.8 and 34.5 (-CH2-), 3.62 (m, 1H, -CH2- at pyrrolidine), 4.37-4.39 (m, 1H, -CH-
44.5 and 44.9 (C-2), 46.7 and 47.1 (-CH2- at pyrrolidine), 59.4 at pyrrolidine), 4.82-4.86 and 4.93-4.94 (m, 1H, H-5), 5.12
and 60.0 (-CH- at pyrrolidine), 67.1 (-C(=O)OCH2Ph), 74.4 (m, 2H, -C(=O)OCH2Ph), 7.29-7.38 (m, 5H, Ph), 7.65 (br s,
and 74.8 (C-5), 127.5 (Ph), 127.8 (Ph), 128.1 and 128.2 (Ph), 1H, NH), 7.93 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3):
128.6 and 128.7 (Ph), 136.5 and 136.6 (Ph), 155.1 (-NC(=O) (rotamer observed) δ 14.2 (C-13), 22.7 (-CH2-), 23.6 and 24.4
OCH2Ph), 172.5 and 172.7 (-OC(=O)CH-), 210.3 and 210.5 (-CH2-), 24.2 and 24.5 (-CH2- at pyrrolidine), 25.3 and 25.4
(C-1). HRMS (FD). Calcd for C25H38N2O4S (M): m/z 462.2552. (-CH2-), 29.2 and 29.3 (-CH2-), 29.4 and 29.5 (-CH2-), 30.3 and
Found: m/z 462.2533.
31.1 (-CH2- at pyrrolidine), 31.9 (-CH2-), 32.8 and 32.9 (-CH2-),
(5R)-5-[(2S)-N-(Benzyloxy)carbonyl-2-pyrrolidi- 33.8 and 34.5 (-CH2-), 44.5 and 44.9 (C-2), 46.7 and 47.1
nylmethanoyloxy]tridecanethioamide [(5R,2’S)-6d1st] (-CH2- at pyrrolidine), 59.4 and 60.0 (-CH- at pyrrolidine),
Yield: 48%; Colorless oil; [α]19D= -33.0 (c= 0.2, MeOH, 67.1 (-C(=O)OCH2Ph), 74.4 and 74.8 (C-5), 127.5 (Ph), 127.8
99% d.e.). Rf= 0.16 (eluent; n-hexane-EtOAc, 3:1, v/v). IR (Ph), 128.1 and 128.2 (Ph), 128.5 and 128.6 (Ph), 136.5 and
(NaCl): cm-1 3312 (N-H), 3203 (N-H), 2929 (CH2, C-H), 2856 136.6 (Ph), 154.6 and 155.1 (-C(=O)OCH2Ph), 172.6 and 172.7
(CH2, C-H), 1739 (OC=O), 1693 (NC(=O)O), 1418 (NHC=S), (-OC(=O)CH-), 210.2 and 210.4 (C-1). HRMS (FD). Calcd for
1120 (C-C(=O)-O), 696 (Ar, C=C). 1H NMR (600 MHz, CDCl3): C26H40N2O4S (M): m/z 476.2709. Found: m/z 476.2676.
(rotamer observed) δ 0.88 (t, J= 6.9 Hz, 3H, H-13), 1.25-1.29
(5R)-5-[(2S)-N-(Benzyloxy)carbonyl-2-pyrrolidi-
[(5R,2’S)-
(m, 12H, H-7, H-8, H-9, H-10, H-11, H-12), 1.50-1.60 (m, 2H, nylmethanoyloxy]tetradecanethioamide
H-6), 1.64-1.67 (m, 2H, H-4), 1.80-1.86 (m, 2H, H-3), 1.88- 6e1st] Yield: 45%; Colorless oil; [α]20D= -38.6 (c= 0.2, MeOH,
2.00 (m, 2H, -CH2- at pyrrolidine), 2.01-2.06 (m, 1H, -CH2- at 99% d.e.). Rf= 0.14 (eluent; n-hexane-EtOAc, 3:1, v/v). IR
pyrrolidine), 2.22-2.28 (m, 1H, -CH2- at pyrrolidine), 2.44- (NaCl): cm-1 3312 (N-H), 3204 (N-H), 2928 (CH2, C-H), 2856
2.49 and 2.61-2.66 (m, 1H, H-2), 2.52-2.57 and 2.71-2.77 (m, (CH2, C-H), 1740 (OC=O), 1692 (NC(=O)O), 1418 (NHC=S),
1H, H-2), 3.49-3.55 (m, 1H- -CH2- at pyrrolidine), 3.61-3.67 1119 (C-C(=O)-O), 696 (Ar, C=C). 1H NMR (600 MHz, CDCl3):
(m, 1H, -CH2- at pyrrolidine), 4.28-4.30 and 4.35-4.37 (m, (rotamer observed) δ 0.88 (t, J= 6.9 Hz, 3H, H-14), 1.25-1.32
1H, -CH- at pyrrolidine), 4.84 and 4.99 (m, 1H, H-5), 5.05 (m, 14H, H-7, H-8, H-9, H-10, H-11, H-12, H-13), 1.48-1.59 (m,
(d, J= 12.4 Hz, 1H, -C(=O)OCH2Ph), 5.10 (d, J= 12.4 Hz, 1H, 2H, H-6), 1.64-1.68 (m, 2H, H-4), 1.80-1.87 (m, 2H, H-3), 1.89-
-C(=O)OCH2Ph), 7.31-7.38 (m, 5H, Ph), 7.18 and 7.54 (br s, 1H, 1.99 (m, 2H, -CH2- at pyrrolidine), 2.01-2.08 (m, 1H, -CH2- at
NH), 7.66 and 8.34 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): pyrrolidine), 2.22-2.29 (m, 1H, -CH2- at pyrrolidine), 2.45-
(rotamer observed) δ 14.2 (C-13), 22.7 and 23.5 (-CH2-), 24.5 2.49 and 2.61-2.66 (m, 1H, H-2), 2.54-2.58, 2.72-2.77 (m, 1H,
and 24.7 (-CH2- at pyrrolidine), 25.0 (-CH2-), 25.3 and 25.4 H-2), 3.50-3.56 (m, 1H, -CH2- at pyrrolidine), 3.61-3.68 (m,
(-CH2-), 29.2 (-CH2-), 29.4 (-CH2-), 29.5 (-CH2-), 30.1 (-CH2- at 1H, -CH2- at pyrrolidine), 4.29 and 4.99 (m, 1H, H-5), 5.05
pyrrolidine), 31.2 and 31.9 (-CH2-), 33.0 and 33.3 (-CH2-), and 5.17 (d, J= 12.4 Hz, 1H, -C(=O)OCH2Ph), 5.10 and 5.19
34.0 and 35.0 (-CH2-), 44.4 and 44.8 (C-2), 46.8 and 47.1 (d, J= 12.4 Hz, 1H, -C(=O)OCH2Ph), 7.05 (br s, 1H, NH), 7.29-
(-CH2- at pyrrolidine), 59.2 and 59.5 (-CH- at pyrrolidine), 7.38 (m, 5H, Ph), 7.46 and 8.33 (br s, 1H, NH). 13C NMR (150
67.0 and 67.3 (-C(=O)OCH2Ph), 74.1 and 74.7 (C-5), 127.5 MHz, CDCl3): (rotamer observed) δ 14.2 (C-14), 22.7 and 23.5
and 127.7 (Ph), 128.0 and 128.2 (Ph), 128.5 and 128.7 (Ph), (-CH2-), 24.5 and 24.7 (-CH2- at pyrrolidine), 25.1 (-CH2-),
136.4 and 136.6 (Ph), 154.5 and 155.2 (-C(=O)OCH2Ph), 172.8 25.4 and 25.5 (-CH2-), 29.3 (-CH2-), 29.4 (-CH2-), 29.6 (-CH2-),
(-OC(=O)CH-), 210.7 and 210.8 (C-1). HRMS (FD). Calcd for 30.1 (-CH2- at pyrrolidine), 31.1 and 31.9 (-CH2-), 33.3 (-CH2-),
C26H40N2O4S (M): m/z 476.2709. Found: m/z 476.2706.
34.0 and 35.0 (-CH2-), 44.4 and 44.8 (C-2), 46.8 and 47.1

Y. Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties ꢁ
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(-CH2- at pyrrolidine), 59.2 and 59.5 (-CH- at pyrrolidine), yields or 5-hydroxyalkanethioamides (4) with about 70%
67.1 and 67.3 (-C(=O)OCH2Ph), 74.1 and 74.7 (C-5), 127.4 and yields. The data for each spectrum was reported above.
127.8 (Ph), 128.0 and 128.2 (Ph), 128.6 and 128.7 (Ph), 136.4
(Ph), 155.2 (-NC(=O)OCH2Ph), 172.8 and 173.1 (-OC(=O)
CH-), 210.9 (C-1). HRMS (FD). Calcd for C27H42N2O4S (M): General procedure for acetylation of optically active
3 and 4
m/z 490.2865. Found: m/z 490.2879.
(5S)-5-[(2S)-N-(Benzyloxy)carbonyl-2-pyrrolidinyl-
methanoyloxy]tetradecanethioamide [(5S,2’S)-6e2nd
]
Acetic anhydride (1.0 mmol, 0.10 g) and 4-dimethyla-
Yield: 48%; Colorless oil; [α]20D= -41.9 (c= 0.2, MeOH, minopyridine (0.1 mmol, 0.01 g) were added to a stirred
99% d.e.). Rf= 0.09 (eluent; n-hexane-EtOAc, 3:1, v/v). IR solution of each of optically active 3 or 4 in anhydrous
(NaCl): cm-1 3309 (N-H), 3204 (N-H), 2928 (CH2, C-H), 2856 CH2Cl2 (5 mL) at room temperature. After 24 h, CH2Cl2 was
(CH2, C-H), 1739 (OC=O), 1693 (NC(=O)O), 1416 (NHC=S), removed under reduced pressure. Distilled water (10 mL)
1118 (C-C(=O)-O), 696 (Ar, C=C). 1H NMR (600 MHz, CDCl3): was then added, and the solution was neutralized with
(rotamer observed) δ 0.88 (t, J= 6.9 Hz, 3H, H-14), 1.21-1.32 Na2CO3. CH3Cl was added to the mixture and the organic
(m, 14H, H-7, H-8, H-9, H-10, H-11, H-12, H-13), 1.44-1.76 layer was separated, washed with distilled water, dried
(m, 6H, H-3, H-4, H-6), 1.88-2.00 (m, 2H, -CH2- at pyrroli- over MgSO4, filtered, and concentrated. Purification of
dine), 2.02-2.07 (m, 1H, -CH2- at pyrrolidine), 2.22-2.31 (m, the crude product by silica gel column chromatography
1H, -CH2- at pyrrolidine), 2.46-2.51 (m, 1H, H-2), 2.53-2.58 (eluent; n-hexane/EtOAc= 3/1) gave the optically active 1
and 2.64-2.69 (m, 1H, H-2), 3.50-3.56 (m, 1H, -CH2- at pyr- and 2 with over 95% yields.
rolidine), 3.59-3.63 (m, 1H, -CH2- at pyrrolidine), 4.36-4.39
(m, 1H, -CH-, at pyrrolidine), 4.82-4.86 and 4.92-4.96 (m,
1H, H-5), 5.12 (m, 2H, -C(=O)OCH2Ph), 7.28-7.38 (m, 5H, Ph), General procedure for the preparation of (R)- or (S)-
7.52 (br s, 1H, NH), 7.91 (br s, 1H, NH). 13C NMR (150 MHz,
CDCl3): (rotamer observed) δ 14.2 (C-14), 22.4 (-CH2-), 24.2
MPA ester
and 24.4 (-CH2-), 23.8 and 24.5 (-CH2- at pyrrolidine), 25.3 A mixture of the resolved (R)-3e (55 mg, 0.2 mmol) or
and 25.4 (-CH2-), 29.3 (-CH2-), 29.5 (-CH2-), 29.6 (-CH2-), 30.3 (R)-4e (52 mg, 0.2 mmol), EDCI (62 mg, 0.4 mmol), DMAP
and 31.1 (-CH2- at pyrrolidine), 32.0 (-CH2-), 32.8 and 32.9 (24 mg, 0.2 mmol) and (R)- or (S)-α-methoxyphenylacetic
(-CH2-), 33.8 and 34.5 (-CH2-), 44.5 and 44.9 (C-2), 46.7 and acid (MPA) (66 mg, 0.4 mmol) in anhydrous THF (10 mL)
47.1 (-CH2- at pyrrolidine), 59.4 and 60.0 (-CH- at pyrroli- was stirred for 2 h at 0 °C, and then stirred for 3 h at room
dine), 67.1 (-C(=O)OCH2Ph), 74.4 and 74.8 (C-5), 127.5 (Ph), temperature. The reaction was quenched with aqueous
127.8 (Ph), 128.1 and 128.2 (Ph), 128.6 and 128.7 (Ph), 136.5 0.1 M HCl (5 mL). The resultant mixture was extracted
and 136.6 (Ph), 154.7 and 155.1 (-C(=O)OCH2Ph), 172.7 and with EtOAc, and the combined extracts were washed with
172.5 (-OC(=O)CH-), 210.3 and 210.5 (C-1). HRMS (FD). Calcd brine, dried over MgSO4, filtered, and concentrated. The
for C27H42N2O4S (M): m/z 490.2865. Found: m/z 490.2868.
crude product was purified by silica gel column chro-
matography (eluent; n-hexane/EtOAc= 3/1) to afford the
corresponding MPA ester [(5R,2’R)- or (5R,2’S)-7e, 8e],
respectively.
(5R)-N-Methyl-5-[(2R)-2-methoxy-2-phenyletha-
noyloxy]tetradecanethioamide [(5R,2’R)-7e] Yield:
General procedure for deacylation of diastereomeric
esters (5 and 6)
To a solution of 5 (1.0 mmol) or 6 (1.0 mmol) in metha- 89%; Colorless oil; [α]22D= -36.5 (c= 0.3, MeOH, 99% d.e.).
nol (1.0 mL) was added a solution of sodium methoxide in Rf= 0.17 (eluent; n-hexane-EtOAc, 3:1, v/v). IR (NaCl): cm-1
methanol (5.0 M, 10 mL). The reaction mixture was stirred 3314 (N-H), 2926 (CH2, C-H), 2854 (CH2, C-H), 1732 (OC=O),
1
for 3 h at 0 °C for the deacylation of 5, and for 30 min at 1456 (Ar, C=C), 697 (Ar, C-H). H NMR (600 MHz, CDCl3):
–15 °C for the deacylation of 6. The reaction was quenched δ 0.88 (t, J= 6.9 Hz, 3H, H-14), 1.22-1.25 (m, 12H, H-7, H-8,
by adding H2O (30 mL) and CHCl3 (10 mL). The resultant H-9, H-10, H-11, H-12), 1.28-1.31 (m, 2H, H-13), 1.34-1.42 (m,
mixture was extracted with CHCl3, and the combined ext- 1H, H-3), 1.43-1.52 (m, 3H, H-3, H-4), 1.53-1.58 (m, 2H, H-6),
racts were washed with brine, dried over MgSO4, filtered 2.47 (m, 2H, H-2), 3.09 (d, J= 4.8 Hz, 3H, CH3NHC(=S)-),
and concentrated. Purification of the crude product by 3.41 (s, 3H, -OCH3), 4.76 (s, 1H, -OC(=O)CH(OCH3)Ph), 4.92
silica gel column chromatography (eluent; n-hexane/ (m, 1H, H-5), 7.32-7.38 (m, 3H, Ph), 7.44-7.45 (m, 2H, Ph),
EtOAc= 1/1) gave the corresponding optically active 7.47 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): δ 14.2 (C-14),
N-methyl-5-hydroxyalkanethioamides (3) with over 90% 22.7 (C-13), 24.7 (C-3, C-4), 25.3 (-CH2-), 29.3 (-CH2-), 29.4

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Y.Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties
(-CH2-), 29.5 (-CH2-), 31.9 (-CH2-), 32.5 (CH3NHC(=S)-), 32.9 (NHC=S), 1494 (Ar, C=C), 1456 (C-N), 1377 (Ar, C=C), 1193
1
(-CH2-), 34.2 (C-6), 45.7 (C-2), 57.3 (-OCH3), 75.1 (C-5), 82.8 (C-C(=O)-O), 734 (Ar, C-H), 698 (Ar, C=C). H NMR (600
(-OC(=O)CH(OCH3)Ph), 127.2 (Ph), 128.7 (Ph), 128.8 (Ph), MHz, CDCl3): δ 0.88 (t, J= 6.9 Hz, 3H, H-14), 0.92 (m, 2H,
136.5 (Ph), 170.9 (-OC(=O)CH(OCH3)Ph), 205.6 (C-1). HRMS H-7), 1.06 (m, 4H, H-8, H-10), 1.13 (m, 2H, H-11), 1.22 (m,
(FD). Calcd for C24H39NO3S (M): m/z 421.2651. Found: m/z 4H, H-9, H-12), 1.28 (m, 2H, H-13), 1.42 (m, 2H, H-6), 1.60 (m,
421.2677.
2H, H-4), 1.69 (m, 1H, H-3), 1.78 (m, 1H, H-3), 2.57-2.62 (m,
(5R)-N-Methyl-5-[(2S)-2-methoxy-2-phenylethano- 1H, H-2), 2.64-2.69 (m, 1H, H-2), 3.40 (s, 3H, -OCH3), 4.77 (s,
yloxy]tetradecanethioamide [(5R,2’S)-7e] Yield: 84%; 1H, -OC(=O)CH(OCH3)Ph), 4.90 (m, 1H, H-5), 7.22 (br s, 1H,
Colorless oil; [α]23D= +45.8 (c= 0.3, MeOH, 99% d.e.). Rf= NH), 7.32-7.37 (m, 3H, Ph), 7.42-7.43 (m, 2H, Ph), 7.66 (br s,
0.15 (eluent; n-hexane-EtOAc, 3:1, v/v). IR (NaCl): cm-1 3312 1H, NH). 13C NMR (150 MHz, CDCl3): δ 14.2 (C-14), 22.7 (C-13),
(N-H), 2925 (CH2, C-H), 2854 (CH2, C-H), 1731 (OC=O), 1455 24.9 (C-3), 25.0 (C-7), 29.2 (-CH2-), 29.3 (-CH2-), 29.4 (-CH2-),
(Ar, C=C), 697 (Ar, C-H). 1H NMR (600 MHz, CDCl3): δ 0.88 29.5 (C-11), 31.9 (-CH2-), 33.1 (C-4), 34.1 (C-6), 44.5 (C-2), 57.3
(t, J= 6.9 Hz, 3H, H-14), 0.92 (m, 2H, H-7), 1.06 (m, 4H, H-8, (-OCH3), 74.8 (C-5), 82.7 (-OC(=O)CH(OCH3)Ph), 127.2 (Ph),
H-10), 1.13 (m, 2H, H-11), 1.22 (m, 4H, H-9, H-12), 1.28 (m, 128.7 (Ph), 128.9 (Ph), 136.4 (Ph), 171.2 (-OC(=O)CH(OCH3)
2H, H-13), 1.42 (m, 2H, H-6), 1.57 (m, 2H, H-4), 1.68 (m, 1H, Ph), 210.3 (C-1). HRMS (FD). Calcd for C23H37NO3S (M): m/z
H-3), 1.80 (m, 1H, H-3), 2.60 (m, 1H, H-2), 2.66 (m, 1H, H-2), 407.2494. Found: m/z 407.2444.
3.12 (d, J= 4.8 Hz, 3H, CH3NHC(=S)-), 3.40 (s, 3H, -OCH3),
4.76 (s, 1H, -OC(=O)CH(OCH3)Ph), 4.89 (m, 1H, H-5), 7.32-
7.37 (m, 3H, Ph), 7.42-7.43 (m, 2H, Ph), 7.72 (br s, 1H, NH). 13C Specific rotation of optically active 5-acetoxy- and 5-hyd-
roxyalkanethioamide analogues
NMR (150 MHz, CDCl3): δ 14.2 (C-14), 22.7 (C-13), 25.0 (C-7),
25.1 (C-3), 29.2 (-CH2-), 29.3 (-CH2-), 29.4 (-CH2-), 29.5 (C-11),
31.9 (-CH2-), 33.0 (C-4), 33.1 (CH3NHC(=S)-), 34.1 (C-6), 45.8 N-Methyl-5-acetoxydodecanethioamide (1c): [α]20D= +8.3 [c
(C-2), 57.3 (-OCH3), 74.9 (C-5), 82.7 (-OC(=O)CH(OCH3)Ph), 0.2, MeOH for (R)-1c], [α]20D= -5.3 [c 0.2, MeOH for (S)-1c].
127.7 (Ph), 128.7 (Ph), 128.8 (Ph), 136.4 (Ph), 171.2 (-OC(=O) N-Methyl-5-acetoxytridecanethioamide (1d): [α]20D= +9.7 [c
CH(OCH3)Ph), 205.7 (C-1). HRMS (FD). Calcd for C24H39NO3S 2.0, MeOH for (R)-1d], [α]20D= -6.7 [c 2.0, MeOH for (S)-1d].
(M): m/z 421.2651. Found: m/z 421.2677.
5-Acetoxydodecanethioamide (2c): [α]25D= +2.4 [c= 0.2,
(5R)-5-[(2R)-2-Methoxy-2-phenylethanoyloxy]tet- MeOH for (R)-2c], [α]25D= -2.5 [c= 0.2, MeOH for (S)-2c].
radecanethioamide [(5R,2’R)-8e] Yield: 61%; Colorless 5-Acetoxytridecanethioamide (2d): [α]20D= +5.7 [c 0.2,
oil; [α]24D= -40.5 (c= 0.3, MeOH, 99% d.e.). Rf= 0.10 (eluent; MeOH for (R)-2d], [α]20D= -8.5 [c 0.2, MeOH for (S)-2d].
n-hexane-EtOAc, 3:1, v/v). IR (NaCl): cm-1 3313 (N-H), 3174 5-Acetoxytetradecanethioamide (2e): [α]20D= +3.9 [c 0.2,
(N-H), 3036 (Ar, C-H), 2927 (CH2, C-H), 2856 (CH2, C-H), MeOH for (R)-2e], [α]20D= -2.1 [c 0.2, MeOH for (S)-2e].
1738 (OC=O), 1626 (NHC=S), 1491 (Ar, C=C), 1457 (C-N), N-Methyl-5-hydroxydodecanethioamide (3c): [α]20D= +2.6
1378 (Ar, C=C), 1179 (C-C(=O)-O), 737 (Ar, C-H), 699 (Ar, [c 0.2, MeOH for (R)-3c], [α]20D= +3.3 [c 0.2, MeOH for (S)-3c].
C=C). H NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.9 Hz, 3H, N-Methyl-5-hydroxytridecanethioamide (3d): [α]20D= +6.3
1
H-14), 1.23-1.25 (m, 12H, H-7, H-8, H-9. H-10, H-11, H-12), 1.28- [c 0.2, MeOH for (R)-3d], [α]20D= -5.2 [c 0.2, MeOH for (S)-3d].
1.32 (m, 2H, H-13), 1.34-1.41 (m, 1H, H-3), 1.43- 1.51 (m, 3H, 5-Hydroxydodecanethioamide (4c): [α]20D= -5.2 [c 0.2,
H-3, H-4), 1.54-1.59 (m, 2H, H-6), 2.41-2.51 (m, 2H, H-2), 3.40 MeOH for (R)-4c], [α]20D= +5.0 [c 0.2, MeOH for (S)-4c].
(s, 3H, -OCH3), 4.76 (m, 1H, -OC(=O)CH(OCH3)Ph), 4.93 (m, 5-Hydroxytridecanethioamide (4d): [α]20D= -1.7 [c 0.2,
1H, H-5), 6.92 (br s, 1H, NH), 7.32-7.38 (m, 3H, Ph), 7.44-7.45 MeOH for (R)-4d], [α]20D= +2.4 [c 0.2, MeOH for (S)-4d].
(m, 2H, Ph), 7.58 (br s, 1H, NH). 13C NMR (150 MHz, CDCl3): 5-Hydroxytetradecanethioamide (4e): [α]20D= -3.4 [c 0.2,
δ 14.2 (C-14), 22.7 (C-13), 24.4 (C-3, C-4), 25.3 (-CH2-), 29.3 MeOH for (R)-4e], [α]20D= +3.9 [c 0.2, MeOH for (S)-4e].
(-CH2-), 29.4 (-CH2-), 29.5 (-CH2-), 31.9 (-CH2-), 32.6 (-CH2-),
34.2 (C-6), 44.4 (C-2), 57.3 (-OCH3), 75.0 (C-5), 82.8 (-OC(=O)
CH(OCH3)Ph), 127.3 (Ph), 128.8 (Ph), 128.9 (Ph), 136.5 (Ph), Lipase screening for enzyme-catalyzed lactonization of
3 and 4
170.9 (-OC(=O)CH(OCH3)Ph), 210.2 (C-1). HRMS (FD). Calcd
for C23H37NO3S (M): m/z 407.2494. Found: m/z 407.2451.
(5R)-5-[(2S)-2-Methoxy-2-phenylethanoyloxy]tet- Novozym 435 (0.4 g, from Candida antarctica B) or other
radecanethioamide [(5R,2’S)-8e] Yield: 66%; Colorless lipase (0.5 w/w, AK from Pseudomonas fluorescens, AS
oil; [α]24D= +44.7 (c= 0.2, MeOH, 99% d.e.). Rf= 0.08 (eluent; from Aspergillus niger, AYS from Candida cylindracea,
n-hexane-EtOAc, 3:1, v/v). IR (NaCl): cm-1 3313 (N-H), 3201 F-AP 15 from Rhizopus oryzae, G from Penicillium camem-
(N-H), 2925 (CH2, C-H), 2856 (CH2, C-H), 1732 (OC=O), 1628 berti, M from Mucor javanicus, PS from Burkholderia

Y. Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties ꢁ
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cepacia, PPL from porcine pancreas) was added to a solu- (C-6), 80.6 (C-5), 172.0 (C-1). HRMS (FI) calcd. for C14H26O2
tion of each substrate (1.0 mmol) in Et2O (20 mL), and the (M)+, 226.1933; found: (M)+, 226.1938.
mixture was stirred at 40 °C for 48 h. After stirring, the
lipase was filtered off and the crude product was purified
by flash chromatography (eluent; n-hexane/EtOAc= 4:1). Antibacterial activity test
All lactones obtained did not show enantioselectivities,
which was confirmed by chiral GC.
The antibacterial assay was performed according to the
δ-Dodecalactone (9c) Colorless oil; Rf= 0.85 (eluent; protocol of the Clinical and Laboratory Standards Insti-
n-hexane-EtOAc, 1:1, v/v). GC conditions [9]: CycloSil-B (30 tute (Wayne, PA, USA). In the protocol, Escherichia coli
m × 0.25 mm i.d. 0.25 μm film thickness) column; oven ATCC25922 and Staphylococcus aureus ATCC29213 are
temperature, 130 °C (isothermal), injector temperature, defined as the quality control strains and were therefore
280 °C, detector temperature, 290 °C, He flow rate : 2.0 used for our antibacterial assay. In the assay, 10 μL of a
mL/min, tR= 73.2 min, tS= 74.4 min. IR (NaCl): cm-1 2953 bacterial cell suspension containing 1×105 colony forming
(CH3), 2927 (CH2), 2872 (CH3), 2857 (CH2), 1738 (OC=O), 1240 units (CFU) bacteria was inoculated into 1 mL of Mueller-
(C-C(=O)-O). 1H NMR (600 MHz, CDCl3): δ 0.88 (t, J= 6.8 Hz, Hinton broth (Becton Dickinson and Company, Sparks,
3H, H-12), 1.22-1.40 (m, 8H, H-8, H-9, H-10, H-11), 1.42-1.62 MD, USA) supplemented with 0.5 mg/mL of the test com-
(m, 4H, H-6, H-7), 1.68 (m, 1H, H-4), 1.78-1.96 (m, 3H, H-3, pound. The culture was incubated for 20 h at 35 °C under
H-4), 2.38-2.62 (m, 2H, H-2), 4.28 (m, 1H, H-5). 13C NMR aerobic conditions, and the viable cells in the culture were
(150 MHz, CDCl3): δ 14.1 (C-12), 18.5 (C-3), 22.6 (-CH2-), 24.9 counted with the modified Miles and Misra method [36],
(-CH2-), 27.8 (C-10), 29.2 (C-11), 29.4 (-CH2-), 29.5 (C-4), 31.8 10 μL aliquots of serial-diluted bacterial suspensions were
(C-6), 35.9 (C-2), 80.6 (C-5), 172.0 (C-1). HRMS (FI) calcd. for inoculated into a trypticase soy agar broth. After 20 h
C12H22O2 (M)+, 198.1620; found: (M)+, 198.1612.
incubation at 35 °C under aerobic conditions, the numbers
δ-Tridecalactone (9d) Colorless oil; Rf= 0.87 (eluent; of colonies recovered from the diluted suspensions on the
n-hexane-EtOAc, 1:1, v/v). GC conditions [11]: CycloSil-B agar broth were counted, and the number of viable bacte-
(30 m × 0.25 mm i.d. 0.25 μm film thickness) column; oven rial cells in the source culture was determined. The anti-
temperature, 130 °C (isothermal), injector temperature, bacterial activity value is defined as follows:
280 °C, detector temperature, 290°C, He flow rate: 2.0
viable cell count of
nagative control [CFU/mL]
mL/min, tR= 124.7 min, tS= 126.9 min. IR (NaCl): cm-1 2953
Antibacterial
(CH3), 2926 (CH2), 2872 (CH3), 2855 (CH2), 1734 (OC=O), 1241
activity value [–] = log
1
viable cell count of
sample[CUF/mL]
(C-C(=O)-O). H NMR (600 MHz, CDCl3): δ 0.88 (t, J= 7.0
Hz, 3H, H-13), 1.22-1.34 (m, 10H, H-8, H-9, H-10, H-11, H-12),
1.42-1.62 (m, 4H, H-6, H-7), 1.68 (m, 1H, H-4), 1.79-1.94 (m,
An antibacterial activity value of >2.0 (≥99% killing
ratio) of a sample treated with synthetic compounds
might be considered as “Antibacterial agent”.
3H, H-3, H-4), 2.40-2.62 (m, 2H, H-2), 4.28 (m, 1H, H-5). 13
C
NMR (150 MHz, CDCl3): δ 14.1 (C-13), 18.5 (C-3), 22.6 (-CH2-),
24.9 (-CH2-), 27.8 (-CH2-), 29.2 (C-11), 29.3 (C-12), 29.4 (-CH2-),
29.5 (C-4), 31.8 (C-6), 35.8 (C-2), 80.6 (C-5), 172.0 (C-1). HRMS
(FI) calcd. for C13H24O2 (M)+, 212.1776; found: (M)+, 212.1776.
δ-Tetradecalactone (9e) Colorless oil; Rf= 0.88 References
(eluent; n-hexane-EtOAc, 1:1, v/v). GC conditions [11]:
[1] Hota S, Hirji Z, Stockton K, Lemieux C, Dedier H, Wolfaaedt
CycloSil-B (30 m × 0.25 mm i.d. 0.25 μm film thickness)
column; oven temperature, 140 °C (isothermal), injector
temperature, 280 °C, detector temperature, 290 °C, He
flow rate: 2.0 mL/min, tR= 118.8 min, tS= 120.8 min. IR
(NaCl): cm-1 2954 (CH3), 2925 (CH2), 2870 (CH3), 2855 (CH2),
G, et al. Outbreak of multidrug-resistant Pseudomonas
aeruginosa colonization and infection secondary to imperfect
intensive care unit room design. Infect. Cont. Hosp. Epdemiol.
2009;30:25–33.
[2] Spellberg B, Guidos R, Gilbert D, Bradley J, Boucher HW,
Scheld WM, et al. The epidemic of antibiotic-resistant
infections: A call to action for the medical community from
the infectious diseases society of America. Clin Infect Dis.
2008;62(2):155–64.
[3] Diekema DJ, Pfaller MA, Schmitz FJ, Smayevsky J, Bell J, Jones
RN, et al. Survey of infections due to Staphylococcus species:
frequency of occurrence and antimicrobial susceptibility of
isolates collected in the United States, Canada, Latin America,
1
1734 (OC=O), 1248 (C-C(=O)-O). H NMR (600 MHz, CDCl3)
δ 0.88 (t, J= 6.8 Hz, 3H, H-14), 1.22-1.34 (m, 12H, H-8, H-9,
H-10, H-11, H-12, H-13), 1.45-1.62 (m, 4H, H-6, H-7), 1.70 (m,
1H, H-4), 1.77-1.96 (m, 3H, H-3, H-4), 2.38-2.62 (m, 2H, H-2),
4.28 (m, 1H, H-5). 13C NMR (150 MHz, CDCl3) δ 14.1 (C-14),
18.5 (C-3), 22.7 (-CH2-), 24.9 (-CH2-×2), 27.8 (-CH2-), 29.3
(-CH2-), 29.4 (C-12), 29.5 (C-13), 29.5 (-CH2-), 31.9 (C-4), 35.9

ꢁ
98ꢁ
Y.Shimotori et al.: Synthesis, antibacterial activities, and sustained perfume release properties
Europe, and the Western Pacific Region for the SENTARY
antimicrobial surveillance program, 1997-1999. Clin Infect Dis.
2001;32:S114–32.
against Gram-positive multiresistant pathogens. Eur J Med
Chem. 2013;65:533–45.
[20] Xue T, Ding S, Guo B, Chu W, Wang H, Yang Y. Synthesis
and structure-relationship studies of novel [6,6,5] tricyclic
oxazolidinone derivatives as potential antibacterial agents.
Bioorg Med Chem Lett. 2015;25:2203–10.
[21] Yamauchi S, Isozaki Y, Nishiwaki H, Akiyama K. Syntheses of
natural 1,3-polyl/α-pyrone and its all stereoisomers to estimate
antifungal activities against plant pathogenic fungi. Bioorg
Med Chem Lett. 2015;25:2189–92.
[22] Du E, Dong W, Guan B, Pan X, Yan Z, Li L, et al. Asymmetric total
synthesis of three stereoisomers of (-)-renieramycin G and
their cytotoxic activities. Tetrahedron. 2015;71:4296–303.
[23] Matsumoto T, Kojima N, Akatsuka A, Yamori T, Dan S, Iwasaki
H, et al. Convergent synthesis of stereoisomers of THF ring
moiety of acetogenin thiophene analogue and their antiproli-
ferative activities against human cancer cell lines. Tetrahedron.
2017;73:2359–66.
[4] Wright GD. Molecular mechanisms of antibiotic resistance.
Chem Commun (Camb). 2011;47:4055–61.
[5] Liu SQ, Yang C, Huang Y, Ding X, Li Y, Fan WM, et al. Antimi-
crobial and antifouling hydrogels formed in situ from
polycarbonate and poly(ethylene glycol) via Michael addition.
Adv Mater. 2012;24:6484–9.
[6] Ramalingam B, Parandhaman T, Das SK. Antibacterial effects of
biosynthesized silver nanoparticles on surface ultrastructure
and nanomechanical properties of gram-negative bacteria viz.
Escherichia coli and Pseudomonas aeruginosa. ACS Appl Mater
Interfaces. 2016;8(7):4963–76.
[7] Singh G, Singh J, Singh A, Singh J, Kumar M, Gupta K, et al.
Synthesis, characterization and antibacterial studies of Schiff
based 1,2,3-triazole bridged silatranes. J Organomet Chem.
2018;871:21–7.
[8] Mustafa YF. Synthesis, characterization and antibacterial
activity of novel heterocycle, coumacine, and two of its
derivatives. Saudi Pharm J. 2018;26:870–5.
[9] Shimotori Y, Hoshi M, Seki S, Osanai T, Okabe H, Ikeda Y, et al.
Preparation of optically pure δ-lactones using diastereomeric
resolution with amino acid as resolving agent. J Oleo Sci.
2015;64(1):75–90.
[10] Shimotori Y, Hoshi M, Miyakoshi T. Enantioselective hydrolysis
and amidation for the preparation of optically active
δ-hexadecalactone. J Oleo Sci. 2015;64(5):561–75.
[11] Shimotori Y, Hoshi M, Okabe H, Miyakoshi T. Preparation of
optically active δ-tri- and δ-tetradecalactones by a combination
of Novozym 435-catalyzed enantioselective methanolysis and
amidation. J Oleo Sci. 2015;64(11):1213–26.
[12] Kanamoto T, Nakashima H, Terakubo S, Miyakoshi T, Itabashi S,
Shimotori Y. Patent JP 2013-147447A.
[13] Krinková J, Doležal M, Hartl J, Buchta V, Pour M. Synthesis
and biological activity of 5-alkyl-6-(alkylsulfanyl)- or 5-alkyl-
6-(arylsulfanyl)pyrazine-2-carboxamides and corresponding
thioamides. Farmaco. 2002;57:71–8.
[14] Yu KL, Torri AF, Luo G, Cianci C, Grant-Young K, Danetz
S, et al. Structure-activity relathioships for a series of
thiobenzamide influenza fusion inhibitors derived from
1,3,3-trimethyl-5-hydroxy-cyclohexylmethylamine. Bioorg Med
Chem Lett. 2002;12:3379–82.
[15] Davies DJ, Faust R, Garratt PJ, Marivingt-Mounir C, Davidson K.
The, M.-T.; Sugden, D. Binding affinity and biological activity
of oxygen and sulfur isosteres at melatonin receptors as a
function of their hydrogen bonding capability. Bioorg Chem.
2004;32:1–12.
[24] Pawlak A, Gładkowski W, Mazur M, Henklewska M, Obmińska-
Mrukowicz B, Rapak A. Optically active stereoisomers of
5-(1-iodoethyl)-4-(4′-isopropylphenyl)dihydrofuran-2-one:
the effect of the configuration of stereocenters on apoptosis
induction in canine cancer cell lines. Chem Biol Interact.
2017;261:18–26.
[25] Zhou L, Wang J, Guo R, Lin B, Wang XB, Huang XX, et al.
Discovery of dihydrobenzofuran neolignans from Rubus ideaus
L. with enantioselective anti-Aβ1-42 aggregation activity. Bioorg
Chem. 2018;80:64–9.
[26] Schnermann P, Schieberle P. Evaluation of key odorants in milk
chocolate and cocoa mass by aroma extract dilution analyses.
J Agric Food Chem. 1997;45:867–72.
[27] Bendall JG. Aroma compounds of fresh milk from New Zealand
cows fed different diets. J Agric Food Chem. 2001;49:4825–32.
[28] Heinemann C, Conde-Petit B, Nuessli J, Escher F. Evidence
of starch inclusion complexation with lactones. J Agric Food
Chem. 2001;49:1370–6.
[29] Yamamoto T, Ogura M, Amano A, Adachi K, Hagiwara T,
Kanisawa T. Synthesis and odor of optically active 2-n-hexyl-
and 2-n-heptylcyclopentanone and the corresponding
δ-lactones. Tetrahedron Lett. 2002;43:9081–4.
[30] Lozano PR, Miracle ER, Krause AJ, Drake M, Cadwallader KR.
Effect of cold storage and packaging material on the major
aroma components of sweet cream butter. J Agric Food Chem.
2007;55:7840–6.
[31] Shimotori Y, Sekine K, Miyakoshi T. Asymmetric synthesis
of δ-lactones with lipase catalyst. Flavour Fragrance J.
2007;22:531–9.
[32] Shimotori Y, Hoshi M, Okabe H, Miyakoshi T, Kanamoto
T, Nakashima H. Synthesis, odour characteristics and
antibacterial activities of the stereoisomeric forms of whisky
lactone and its thiono analogues. Flavour Fragrance J.
2017;32:29–35.
[16] Boström J, Olsson RI, Tholander J, Greasley PJ, Ryberg E,
Nordberg H, et al. Novel thioamide derivatives as neutral CB1
receptor antagonists. Bioorg Med Chem Lett. 2010;20:
479–82.
[17] Thanigaimalai P, Sharma VK, Lee KC, Yun CY, Kim Y, Jung SH.
Refinement of the pharmacophore of 3,4-dihydroquinazoline-
2(1H)-thiones for their anti-melanogenesis activity. Bioorg Med
Chem Lett. 2010;20:4771–3.
[18] Brunhofer G, Studenik C, Ecker GF, Erker T. Synthesis,
spasmolytic activity and structure-activity relationship study of
a series of polypharmacological thiobenzanilides. Eur J Pharm
Sci. 2011;42:37–44.
[33] Trost BM, Belletire JL, Godleski S, McDougal PG, Balkovec JM.
On the use of the O-methylmandelate ester for establishment
of absolute configuration of secondary alcohols, 1986, 51,
2370–2374.
[34] Trost BM, Bunt RC, Pulley SR. On the use of O-methylmandelic
acid for the establishment of absolute configuration of α-chiral
primary amines, 1994, 59, 4202–4205.
[35] Nobuhiro A. Patent JP P2003-313580A.
[19] Fortuna CG, Bonaccorso C, Bulbarelli A, Caltabiano G, Rizzi
L, Goracci L, et al. New linezolid-like 1,2,4-oxadiazoles active
[36] Miles AA, Misra SS, Irwin JQ. The estimation of the bactericidal
power of the blood. J Hyg (Lond). 1938 Nov;38:732.