Structure-activity relationships of globomycin analogues as antibiotics

Article abstract of DOI:10.1016/j.bmc.2003.10.055

Globomycin (1a), a signal peptidase II inhibitor, and its derivatives show potent antibacterial activity against Gram-negative bacteria. The synthesis and antimicrobial activity of novel globomycin analogues are reported. The hydroxyl group in the L-Ser r

Full text of DOI:10.1016/j.bmc.2003.10.055

Bioorganic & Medicinal Chemistry 12 (2004) 337–361
Structure–activity relationships of globomycin analogues
as antibiotics
Toshihiro Kiho,a Mizuka Nakayama,a Kayo Yasuda,b Shunichi Miyakoshi,b
Masatoshi Inukaib and Hiroshi Kogena,*
aExploratory Chemistry Research Laboratories, Sankyo Co., Ltd., 2-58, Hiromachi 1-chome, Sinagawa-ku,
Tokyo 140-8710, Japan
bLead Discovery Research Laboratories, Sankyo Co., Ltd., 2-58, Hiromachi 1-chome, Sinagawa-ku, Tokyo 140-8710, Japan
Received 6 September 2003; revised 30 October 2003; accepted 31 October 2003
Abstract—Globomycin (1a), a signal peptidase II inhibitor, and its derivatives show potent antibacterial activity against Gram-
negative bacteria. The synthesis and antimicrobial activity of novel globomycin analogues are reported. The hydroxyl group in the
l-Ser residue was essential for the antimicrobial activity and the length of the alkyl side chain greatly influenced the activity. In
addition, derivatives that had a modified cyclic core exhibited weak activity. One of the analogues showed a wider antimicrobial
spectrum, effective against not only Gram-negative but also Gram-positive bacteria.
# 2003 Elsevier Ltd. All rights reserved.
1. Introduction
geners have the structure shown in Figure 1. The minor
congeners, SF-1902 A3 and A4b, have an l-Val in the
place of l-allo-Ile, and the other congeners, SF-1902 A2,
A4a and A4b, have a shorter or longer alkyl side chain in
the fatty acid unit than that of 1a. It was reported that
the antibacterial activity is quite sensitive to the length
of the alkyl side chain, either in the fatty acid or amino
acid.2 The congeners which have a longer side chain, 1b,
SF-1902 A4a and A4b, are more potent than 1a (MIC:
1a, 6.25 mg/mL; 1b, 1.56 mg/mL against E. coli NIHJ
JC-2). However, SF-1902 A2 with the shortest side chain
showed the weakest activity.2b Furthermore, the con-
geners containing an l-Val, SF-1902 A3 and A4b, are
less active compared with 1a and 1b, respectively. In
addition, based on 1H NMR analysis, 1a exists as a
mixture of rotational isomers derived from the acyl N-
methyl amide group. The major isomer was ascribed to
be in the trans form while the minor isomer was con-
sidered to adopt a cis form.1d,e The cis/trans ratio may
affect the antibacterial activity. Now, we wish to report
the full structure–activity relationships (SARs) of newly
synthesized globomycin analogues.
Globomycin (1a)1 and its congeners, SF-1902 A2–A5,2
isolated by different researchers as antibiotics against
only Gram-negative bacteria, are 19-membered cyclic
depsipeptides.1,2 The major component, 1a, has only
been proven to be a specific inhibitor of signal peptidase
II, a prolipoprotein-processing enzyme,3 that processes
the acylated precursor form of lipoprotein into apoli-
poprotein and a signal peptide in Escherichia coli.4
Inhibition of signal peptidase II leads to the accumula-
tion of the lipoprotein precursor in the cytoplasmic
membrane and consequently to the death of the cell.5
Signal peptidase II represents an attractive target because
the mechanism is different from currently available
drugs. Previously, we reported the absolute structure of
1a obtained by X-ray analysis and the first asymmetric
total synthesis of globomycin (1a) and SF-1902A5
(1b).1d,e In a recent communication, we also reported on
new synthetic globomycin analogues that have more
potent inhibitory activity.1f Structurally, these con-
geners were constructed from four natural amino acids,
one N-methyl amino acid and a b-hydroxy-a-methyl
carboxylic acid. Naturally occurring globomycin con-
2. Chemistry
Three fragments (Fragments A, B and C) were synthe-
sized and assembled to construct various globomycin
* Corresponding author. Tel.: +81-3-3492-3131; fax: +81-3-5436-
8561; e-mail: hkogen@shina.sankyo.co.jp
0968-0896/$ - see front matter # 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmc.2003.10.055

338
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
chiral auxiliary followed by the reduction in the presence
of Boc2O gave (2R,3S)-3-Boc-amino-2-methylnonanoic
acid 9 (46%, two steps). Finally, (3R)-hydroxynonanoic
acid 118 was produced by asymmetric hydrogenation of
b-ketoester 10 with a BINAP ruthenium catalyst9 fol-
lowed by hydrolysis (96%, two steps). The optical pur-
ity was confirmed by 1H NMR analysis after the
conversion to (R)- and (S)-MTPA ester.
The dipeptide unit, Fragment B (15a–15e), was prepared
from commercially available l-allo-threonine (l-allo-
Thr-OH), Boc-l-threonine [Boc-l-Thr-OH (12a)] or
Boc-l-a-aminobutyric acid (Boc-l-Abu-OH). Scheme 3
shows the preparation of compound 15b as an example
and a representative structure of Fragment B. The pro-
tection of 12 with TBSOTf (88%) followed by con-
densation with glycine benzyl ester (13) afforded fully
protected dipeptide 14b (87%). Compound 14b was
hydrogenated to give the dipeptide Fragment B 15b
(98%). In the case of 15c, N-Me-Gly-OBn was used in
place of 13. Dipeptide 15e was also synthesized with
Boc-l-allo-Thr(Me)-OH (12b) prepared from l-allo-
Thr-OH.
Figure 1. Structure of naturally occurring globomycin (1a) and its
congeners.
analogues as shown in Scheme 1. Each fragment was
prepared with the corresponding method used in the
total synthesis of globomycin (1a).1d Fragment A is an
optically active b-hydroxy or b-amino carboxylic acid.
Fragments B and C are dipeptidyl and tripeptidyl com-
pounds, respectively. Fragment A, the only non-amino
acid containing part, was prepared as shown in Scheme
2. Compound 31d,e was prepared by an anti-selective
boron-mediated asymmetric aldol reaction developed by
Abiko and Masamune.6 The anti-aldol product 3c was
obtained in good yield and with high diastereoselectivity
(n=7: 76%, 89% de). The hydroxyl ester 3c was
hydrolyzed with aqueous LiOH in THF to give 4c
without epimerization in good yield (86%). On the
other hand, the syn isomer 7 was prepared by a syn-
selective Evans aldol method7 with a chiral oxazolidi-
none 5. The aldol product 6 (93%) was hydrolyzed to
give an acid 77c in the usual manner (100%). The
hydroxyl groupin 6 was converted to an azide groupby
Mistunobu inversion to afford 8 (81%). Removal of the
The tripeptide 19a–19f, Fragment C, was synthesized
from Boc-protected l-Serine derivatives 16 as shown in
Scheme 4. An allyl or benzyl groupwas selected to
protect the carboxylic acid in 16 (P1=Me, P2=Bn or
P1=Bn, P2=allyl). The esterification of 16 followed by
the deprotection of the Boc group with 4 N HCl in
EtOAc gave the ester 17 as an HCl salt. Coupling of this
salt with Boc-l-allo-isoleucine (Boc-l-allo-Ile-OH)
mediated by (benzotriazolyloxy) tris(pyrrolydino)phos-
phonium hexafluorophosphate (PyBOP)10 was con-
ducted to give dipeptide 18 (100%). After the removal
of the Boc groupin 18, the coupling with a variety of
Boc-protected amino acid followed by the treatment
with 4 N HCl in EtOAc gave tripeptides 19a–19e. Tri-
peptide 19g was also prepared in the same manner but
by using Boc-l-Ile-OH in palce of Boc-l-allo-Ile-OH.
The novel synthetic analogues 1c–1q were prepared by
the convergent macrolactamization method using the
Scheme 1. Synthetic strategy for globomycin analogues.

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
339
three fragments (Fragments A, B and C). As an exam-
ple, synthesis of the most potent analogue 1c is shown in
Scheme 5. Condensation of Fragment C (19b) with
Fragment A (4c) mediated by diethylcyanophosphate
(DEPC)11 was carried out to give the acylated tripeptide
20c (99%). Esterification of 20 was performed with di-
isopropylcarbodiimide (DIPC) and Fragment B (15a)
under Keck’s condition12 to afford depsipeptide 21c
(97%). After the treatment of 21c with TBAF in the
presence of AcOH (91%), the removal of the allyl
(91%) and Boc groupwith a Pd catalyst and TFA
respectively, provided the macrocyclization precursors.
The macrolactamization was performed with O-(7-
azabenzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexa-
fluorophosphate (HATU)13 to give O-Bn cyclic deriva-
tives under highly diluted conditions (33%, two steps).
Finally, the removal of the benzyl groupby hydro-
genolysis yielded 1c (90%). The other analogues 1d–1q
were also synthesized by the same route.14 The reagent,
2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
tetrafluoroborate (TBTU),13 was also used for macro-
cyclization. The newly obtained globomycin analogues
1
are shown in Figure 2. The H NMR analysis indicated
these analogues were present as a mixture of rotamers,
but the N-demethyl derivative 1j was found to be single
isomer.15
3. Antibacterial activity
The antibacterial activities of the synthetic globomycin
analogues 1a–1q against Gram-negative bacteria are
summarized in Table 1. As a result, 1c1f possessing the
longest alkyl side chain showed the most potent activity
among the analogues (MIC: 1a, 12.5 mg/mL; 1b, 3.13
mg/mL; 1c, 1.56 mg/mL against E. coli SANK 70569).
Scheme 2. Synthesis of Fragment A.
Scheme 3. Synthesis of the dipeptide Fragment B.

340
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
The length of the alkyl side chain greatly affected the
antibacterial activity. A four-carbon increase in the
fatty acid side chain enhanced the activity by 4- to 8-
fold compared with 1a while the cis/trans geometric
ratio was almost constant among these three com-
pounds. The hydrophobicity seemed to be important for
the antibacterial activity, therefore, it may be possible to
produce other more potent inhibitors. With regard to
stereochemistry, the activity of 1d, C(25)-epimer, was
diminished and the activities of 1e, C(22)-epimer, and 1f,
stereoisomer at C(22) and C(25) positions, were com-
pletely lost. In particular, the stereochemistry of the
hydroxyl groupin l-Thr was quite important for the
activity. Compound 1e was inactive although the deoxy
derivative 1g and methyl ether derivative 1h retained their
activity. Therefore, the hydroxyl groupin l-allo-Thr is
Scheme 4. Synthesis of the tripeptide Fragment C.
Scheme 5. Synthesis of the globomycin derivative 1c. Reagents and conditions: (a) DEPC, Et3N,THF, 0ꢀC to rt, 99%; (b) DIPC, CSA, DMAP,
CH2Cl2, 0ꢀC to rt, 97%; (c) TBAF, AcOH, 91%; (d) cat Pd(PPh3)4, morpholine, 94%; (e) TFA, CH2Cl2; (f) HATU or TBTU, i-Pr2NEt, 33% (two
steps); (g) H2, Pd(OH)2, 90%.

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
341
Figure 2. Structures of globomycin (1a) and the synthetic analogues (1b–1q).
1
not essential for the activity. On the other hand, the
hydroxyl groupin l-Ser is essential because generating
a methyl ether derivative, such as 1i, lost the activity.16
Among derivatives with a modified N-alkyl moiety, 1i–
1m, only N-ethyl derivative 1k showed weak activity.
However the other derivatives, 1j, 1l and 1m did not
(MIC: 1k, 25 mg/mL against E. coli SANK 70569). H
NMR analysis of these compounds suggested that the
conformation of 1k was similar to that of globomycin
(1a)17 but the conformations of 1j, 1l and 1m were quite
different. Furthermore, the NOESY experiments of
these compounds indicated that conformation of the
major isomer at the acyl N-H or N-alkyl amide moiety
was trans form which is the same as 1a, as shown in
Figure 3. These results suggested that modifications at
the N-Me position caused a conformational change at
another site in the molecule. Taking this into account,
lactam 1o, syn 1p and N-Me-Gly derivative 1q were
inactive because of a conformational change ascribed to
a modification of the cyclic core.
Surprisingly, 1c showed moderate activity against all
Gram-positive bacteria tested such as Staphylococcus
aureus (MRSA) (MIC=12.5 mg/mL) even though 1a
and 1b were almost inactive as shown in Table 2. This is
the first globomycin analogue that has a wide anti-
Figure 3. Selected NOESY Structure of 1l and the ratio of rotamers in
CDCl3.

342
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
Table 1. Antibacterial spectra of globomycin (1a), SF-1902 A5 (1b) and the synthetic analogues (1c–1i)
Compd
MIC (mg/mL)
Organism
Escherichia
coli
Escherichia
coli
Salmonella
enteritidis
Klebsiella
pneumoniae
Enterobacter
cloacae
846
Enterobacter
cloacae
Serratia
marcescens
Pseudomonas
aeruginosa
SANK 72690 SANK 72790 SANK 73190
SANK 70569 SANK 72290 SANK 72390 SANK 72490
(NIHJ JC-2)
Ampicillin
Streptomycin,
50
12.5
>100
>100
12.5
3.13
>100
6.25
100
6.25
>100
3.13
50
3.13
>100
50
sulfate
1a
1b
1c
1d
1e
12.5
12.5
3.13
1.56
12.5
>100
>100
25
25
>100
>100
25
>100
>100
>100
>100
>100
>100
25
6.25
3.13
50
25
3.13
1.56
25
6.25
1.56
1.56
6.25
50
12.5
3.13
100
12.5
3.13
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
3.13
1.56
12.5
>100
>100
25
25
>100
>100
25
>100
>100
>100
>100
>100
>100
50
>100
>100
>100
100
>100
>100
100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
100
>100
>100
50
100
1f
>100
12.5
12.5
>100
100
1g
1h
1i
1j
1k
1l
1m
1n
1o
1p
1q
50
50
>100
>100
50
>100
>100
>100
>100
>100
>100
>100
>100
50
>100
>100
>100
>100
>100
>100
12.5
>100
>100
>100
>100
>100
>100
bacterial spectrum which also includes Gram-positive
bacteria. These results suggest that lipoproteins are
essential for not only Gram-negative bacteria but also
Gram-positive bacteria and that signal peptidase II
inhibitors would probably be effective against all bac-
teria. Finding such an inhibitor would lead to the
development of a new class of antibiotics. Finally, the
antifungal activity of these analogues was tested. How-
ever, no activity was observed against Candida albicans,
Candida glabrata and Aspergillus clavatus (>50 mg/mL).
(CH2Cl2) was distilled from calcium hydride. Other
anhydrous solvents were purchased from Aldrich or
Kanto Kagaku. All reagents were commercially avail-
able and used as obtained unless otherwise stated. Pre-
parative flash column chromatography was performed
using Merck Silica gel 60 (230–400 mesh). NMR spectra
were obtained on a JEOL ECP-500, JNM-GSX-400,
1
Varian Inova-500, Mercury-400. All H NMR spectra
are reported in ppm downfield from tetramethylsilane as
an internal standard. All 13C NMR spectra are reported
in ppm relative to the central line for CDCl3 (d 77.0) or
CD3OD (d 49.0). The following abbreviations were used
to explain the multiplicities: s=singlet, d=doublet,
t=triplet, q=quartet, qu=quintet, m=multiplet,
br=broadened. In the NMR spectral lists, chemical
shifts which are assigned to the minor conformer are
marked with an asterisk.
In summary, new globomycin analogues were synthe-
sized and their antibacterial activities against Gram-
negative bacteria were evaluated. SAR studies were
conducted based on the NMR spectroscopic analysis of
the analogues. Compound 1c possessing the longest
alkyl side chain showed the most potent activity among
the synthesized analogues and the antibacterial spec-
trum was expanded to also includes Gram-positive bac-
teria. These results were promising in the search to find
a more potent and effective antibiotic.
4.1.1. Fragment A: (2R,3R)-3-hydroxy-2-methyltrideca-
noic acid (1R,2S)-2-[benzyl-(2,4,6-trimethyl-benzenesul-
fonyl)amino]-1-phenylpropyl ester (3c). To a stirred
solution of (1R,2S)-2 (701 mg, 1.46 mmol) in CH2Cl2
(15 mL) was added Et3N (0.52 mL, 3.73 mmol). The
solution was cooled to À78 ꢀC and to this was trans-
ferred via cannula a solution of c-Hex2BOTf (1.0 M in
hexane, 3.05 mL, 3.05 mmol), which was precooled to
À78 ꢀC. The resulting solution was stirred at À78 ꢀC for
2 h to complete enolization. A solution of undecylic
aldehyde (615 mg, 3.61 mmol) in CH2Cl2 (5 mL) was
added dropwise to the enolate solution and the reaction
mixture was stirred at À78 ꢀC for 4 h and at 0 ꢀC for 2 h.
The reaction was quenched by the addition of pH 7
buffer solution (6 mL) followed by CH3OH (15 mL) and
30% H2O2 (1.5 mL). The mixture was stirred overnight
vigorously at room temperature. The resulting solution
was extracted with ether. The organic extracts were
3.1. Measurement of antibacterial activity
Bacteria were inoculated on Nutrient Agar (Eiken Chem-
ical Co., Ltd.) and the MICs of the synthesized analogues
were determined by the agar dilution method.18
4. Experimental
4.1. General procedure
All moisture-sensitive reactions were carried out under a
N2 atmosphere. Tetrahydrofuran (THF) was distilled
from sodium-benzophenone ketyl. Dichloromethane

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
Table 2. Antibacterial spectra of globomycin (1a–1c) against Gram-positive bacteria
343
Compd
MIC (mg/mL)
Organism
Staphylococcus aureus
SANK 70668
Staphylococcus aureus
SANK 71790
Staphylococcus aureus
SANK 71890a
Enterococcus faecalisa
SANK 71990
Ampicillin
Streptomycin, sulfate
1a
1b
1c
0.78
1.56
>100
50
1.56
3.13
>100
50
>100
>100
>100
50
12.5
100
>100
100
6.25
6.25
12.5
12.5
a MRSA.
combined, dried over MgSO4, filtered and concentrated
in vacuo. The resulting crude product was partially
purified by silica gel flash column chromatography with
a 10:1 mixture of hexane and EtOAc as an eluent to
afford a mixture of aldol products (825 mg, 1.27 mmol,
87%, 89% de). The ratio of the diastereomers
(94.67:5.06:0.25) was determined by HPLC analysis
(Daicel Chiralcel AD 0.46 cmÂ25 cm, n-hexane/i-
PrOH=92:8, 1.0 mL/min, 40 ꢀC). Purification of the
diastereomers by silica gel flash column chromato-
graphy with a 10:1 mixture of hexane and EtOAc as an
eluent to give compound 3c as a colorless oil (705 mg,
6H2O: C, 67.97; H, 11.54; Found: C, 68.11; H, 11.79;
[a]2D4 +2.7 (c 1.03, CHCl3).
4.1.3. (4R,5S)-3-[(2R,3S)-(3-Hydroxy-2-methylnonanoyl)]-
4-methyl-5-phenyloxazolidin-2-one (6). A chiral oxazoli-
dione 5 (2.90 g, 12.4 mmol) was dissolved in CH2Cl2 (25
mL) and cooled at 0 ꢀC. To this solution were added n-
Bu2BOTf (1.0 M solution in CH2Cl2, 13.7 mL, 13.7
mmol) and Et3N (2.10 mL, 15.1 mmol) at 0 ꢀC. The The
resulting solution was stirred at 0 ꢀC for 15 min and at-
15 ꢀC for 15 min. After the mixture was cooled to
À78 ꢀC, heptanal (1.56 g, 13.7 mmol) was added drop-
wise to the enolate solution. The reaction mixture was
stirred at À78 ꢀC for 2 h, at À10 ꢀC for 20 min and then
at 0 ꢀC for 2 h. The reaction was quenched by the addi-
tion of aqueous phospahte buffer solution (14 mL) fol-
lowed by CH3OH (40 mL) and 30% H2O2 (15 mL). The
mixture was stirred at 0 ꢀC for 1 h and then evaporated.
The resulting solution was extracted with ether. The
organic extracts were combined, dried over MgSO4, fil-
tered and concentrated in vacuo. The resulting crude
product was purified by silica gel flash column chroma-
tography with a gradient elution system, a 6:1–5:1 mix-
ture of hexane and EtOAc as eluents to give 6 (4.01 g,
11.5 mmol, 93%) as a colorless crystal (mp108–109 ꢀC
). 1H NMR (400 MHz, CDCl3) d ppm: 0.89 (t, 3H,
J=7.0 Hz), 0.90 (d, 3H, J=6.8 Hz), 1.24 (d, 3H, J=7.0
Hz), 1.29–1.59 (m, 10H), 2.88 (d, 1H, J=3.0 Hz) 3.78
(dq, 1H, J=2.7, 7.0 Hz), 3.94–3.99 (m, 1H), 4.80 (qu,
1H, J=6.8 Hz), 5.69 (d, 1H, J=7.3 Hz), 7.30–7.32 (m,
2H), 7.36–7.45 (m, 3H); 13C NMR (100 MHz, CDCl3) d
ppm: 10.2, 14.1, 14.4, 22.6, 26.0, 29.2, 31.8, 33.9,
42.2, 54.8, 71.6, 78.9, 125.6, 128.8, 128.9, 133.2, 152.6,
177.5; IR (KBr) cmÀ1: 3462, 2949, 2922, 1787, 1690,
1370, 1346, 1242, 1195, 1154; HRMS calcd for
C20H30NO4 (M+H)+ calcd 348.2175, found 348.2177.
Anal. calcd for C20H29NO4: C, 69.14; H, 8.41, N, 4.03;
Found: C, 69.43; H, 8.38, N, 4.06; [a]2D5 +18.0 (c 1.14,
CHCl3).
1
1.08 mmol, 76%). H NMR (500 MHz, CDCl3) d ppm:
0.88 (t, 3H, J=6.6 Hz), 1.13 (d, 3H, J=7.3 Hz), 1.18 (d,
3H, J=7.3 Hz), 1.20–1.47 (m, 19H), 2.28 (s, 3H), 2.43–
2.51 (m, 1H); 2.48 (s, 6H), 3.60–3.64 (m, 1H), 4.12 (dq,
1H, J=7.3 Hz), 4.54 and 4.76 (AB type d’s, each 1H,
J=16.9 Hz) 5.83 (d, 1H, J=4.4 Hz) 6.86-6.88 (m, 4H),
7.16–7.30 (m, 8H); 13C NMR (125 MHz, CDCl3) d ppm:
13.5, 14.1, 20.9, 22.66, 22.69, 25.4, 29.3, 29.6, 29.6, 31.6,
31.9, 34.5, 45.5, 48.3, 56.7, 73.2, 78.2, 126.0, 127.2,
127.7, 128.0, 128.3, 128.4, 132.1, 133.4, 138.2, 138.4,
140.3, 142.6, 174.6; IR (CHCl3) cmÀ1: 2982, 2928, 2856,
1729, 1605, 1456, 1382, 1322, 1153, 855; HRMS calcd
for C39H55NO5SNa (M+Na)+ calcd 672.3698, found
672.3716; [a]2D5 +19.7 (c 1.16, CHCl3).
4.1.2. (2R,3R)-3-hydroxy-2-methyltridecanoic acid (4c).
.
A solution of 3c (204 mg, 0.313 mmol) and LiOH H2O
(65.9 mg, 1.57 mmol) in THF–CH3OH–H2O (2:3:2, 7.0
mL) was stirred at room temperature overnight. The
mixture was poured into water (5 mL) and extracted
with CH2Cl2. The aqueous layer was acidified with 10%
aqueous KHSO4 solution and extracted with CH2Cl2.
The extracts were washed with brine, dried with
MgSO4, filtered and evaporated. The residue was pur-
ified by silica gel flash column chromatography with a
gradient elution system, a 3:1–1:0 mixture of hexane and
EtOAc as eluents to give 4c (65.8 mg, 0.269 mmol, 86%)
as a colorless solid (mp44–45 ꢀC). H NMR (400 MHz,
1
CDCl3) d ppm: 0.88 (t, 3H, J=6.9 Hz), 1.25 (d, 3H,
J=7.0 Hz), 1.26–1.50 (m, 16H), 1.50–1.60 (m, 2H), 2.56
(qu, 1H, J=7.0 Hz), 3.68–3.72 (m, 1H); 13C NMR
(125 MHz, CDCl3) d ppm: 14.1, 14.2, 22.7, 25.4, 29.3,
4.1.4. (4R,5S)-3-{(2R,3R)-(3-Azide-2-methylnonanoyl)}-
4-methyl-5-phenyloxazolidin-2-one (8). To a solution of
6 (198 mg, 0.569 mmol) in THF (3.0 mL) were DEAD
(99 mL, 0.626 mmol) Ph3P (164 mg, 0.626 mmol) at
room temperature. After being stirred for 5 min, DPPA
(35 mL, 0.626 mmol) was added. This reaction mixture
was stirred over night and evaporated. The residue was
purified by silica gel flash column chromatography with
29.5, 29.6, 31.9, 34.6, 45.2, 73.3, 180.9; IR (KBr) cmÀ1
:
3252, 2916, 2849, 2617, 1710, 1689, 1465, 1283, 1265,
1091; HRMS calcd for C14H29O3 (M+H)+ calcd
.
245.2117, found 245.2121. Anal. calcd for C14H28O3 1/

344
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
a 10:1 mixture of hexane and EtOAc as an eluent to give
8 (171 mg, 0.458 mmol, 81%) as a colorless oil, 1H
NMR (400 MHz, CDCl3) d ppm: 0.87–0.94 (m, 6H),
1.20 (d, 3H, J=6.9 Hz), 1.24–1.55 (m, 9H), 1.66–1.74
(m, 1H), 3.70 (dt, 1H, J=2.8, 9.1 Hz), 3.80–3.88 (m,
1H), 4.83 (qu, 1H, J=6.9 Hz), 5.72 (d, 1H, J=7.2 Hz),
7.31–7.33 (m, 2H), 7.36–7.45 (m, 3H); 13C NMR
(125 MHz, CDCl3) d ppm: 14.1, 14.3, 14.6, 22.6, 25.6,
29.1, 31.3, 31.7, 42.3, 55.1, 64.9, 79.0, 125.6, 128.76,
128.81, 133.2, 152.6, 174.7; IR (CHCl3) cmÀ1: 2930,
2859, 2104, 1781, 1698, 1456, 1386, 1345, 1254, 1121;
HRMS calcd for C20H29N4O3 (M+H)+ 373.2240,
found 373.2249; [a]2D6 À5.12 (c 1.35, CHCl3).
graphy with a gradient elution system, a 10:1–8:1
mixture of hexane and EtOAc as eluents to give methyl
(R)-3-hydroxynonanoate (1.34 g, 7.12 mmol, 96%) as a
1
slightly yellow oil. H NMR (400 MHz, CDCl3) d ppm:
0.88 (t, 3H, J=7.0 Hz), 1.25–1.38 (m, 7H), 1.38–1.56
(m, 3H), 2.41 (dd, 1H, J=8.9, 16.3 Hz) 2.52 (dd, 1H,
J=3.3, 16.3 Hz), 2.87 (br s, 1H), 3.72 (s, 3H), 3.97–4.04
(m, 1H); 13C NMR (125 MHz, CDCl3) d ppm: 14.0,
22.6, 25.4, 29.2, 31.7, 36.5, 41.1, 51.7, 68.0, 173.5; IR
(CHCl3) cmÀ1: 3596, 2956, 2931, 2859, 1724, 1439, 1404,
1335, 1267, 1174; HRMS calcd for C10H21O3 (M+H)+
calcd 189.1491, found 189.1481; [a]2D4À20.0 (c 1.33,
CHCl3).
4.1.5. (2R,3R)-3-Boc-amino-2-methylnonanoic acid (9).
Compound 8 (193 mg, 0.518 mmol) was dissolved in
THF–H2O (4:1.3, 4.3 mL). To this solution were added
To the solution of this ester (557 mg, 2.96 mmol) in
CH3OH (20.0 mL) was added 1 N NaOH aqueous
solution (6.0 mL, 6.0 mmol) at 0 ꢀC. The mixture was
stirred at the same temperature for 30 min and at room
temperature for an additional 3 h. The solution was
acidified with 5% aqueous HCl solution and extracted
with EtOAc. The organic extracts were washed with
brine, dried with MgSO4, filtered and evaporated to give
11 (514 mg, 2.95 mmol, 100%) as a colorless solid (mp
.
LiOH H2O (44.0 mg, 1.04 mmol) and 30% aqueous
ꢀ
H2O2 (0.1 mL) at C. After being stirred at the same
temperature for 2 h, 10% Na2S2O3 aqueous solution
was added. The reaction mixture was acidified with 10%
aqueous KHSO4 solution and extracted with CH2Cl2.
The combined organic layer was dried over Na2SO4,
filtered and evaporated to give (2R,3R)-3-azide-2-
methyltnonanoic acid (83.5 mg, 0.391 mmol, 76%) as a
colorless oil.
49–50 ꢀC). H NMR (400 MHz, CDCl3) d ppm: 0.89 (t,
1
3H, J=7.1 Hz), 1.25–1.40 (m, 7H), 1.40–1.50 (m, 2H),
1.52–1.60 (m, 1H), 2.48 (dd, 1H, J=8.9, 16.3 Hz) 2.58
(dd, 1H, J=3.4, 16.3 Hz), 4.01–4.07 (m, 1H); 13C NMR
(125 MHz, CDCl3) d ppm: 14.0, 22.6, 25.4, 29.1, 31.7,
36.5, 41.0, 68.0, 177.8; IR (KBr) cmÀ1: 3481, 3121, 2956,
2927, 2857, 1740, 1717, 1683, 1414, 1176; HRMS calcd
for C9H19O3 (M+H)+ calcd 175.1334, found 175.1322.
Anal. calcd for C9H18O3: C, 62.04; H, 10.41; Found: C,
61.75; H, 10.20; [a]2D4=À20.5 (c 1.14, CHCl3).
This azide (280 mg, 1.31 mmol) and Boc2O (343 mg,
1.57 mmol) were dissolved in EtOAc (3.0 mL). To this
solution, 10% Pd/C (cat) was added and the resulting
mixture was stirred at room temperature for 10 h under
H2 atmosphere. Pd/C was removed by filtration and the
filtrate was evaporated. The residue was purified by
column chromatography with a gradient elution system,
a 1:0-20:1 mixture of CH2Cl2 and CH3OH as eluents to
give 9 (231 mg, 3.95 mmol, 61%) as a colorless solid
(mp, 84 ꢀC). 1H NMR (400 MHz, CDCl3, two con-
formers (major/minor=3.0/1)) d ppm: 0.86–0.89 (m,
3H), 1.17–1.31 (m, 13H), 1.44 (s, 27/4H), 1.46 (s, 9/4H),
2.63-2.74 (m, 1H), 3.65-3.78 (m, 1H), 5.06 (br d, 3/4H,
J=10.0 Hz), 5.56 (br d, 1/4H, J=9.0 Hz); 13C NMR
(125 MHz, CDCl3, both rotamers) d ppm: 14.1, 14.6,
16.8, 22.56, 22.64, 26.3, 27.2, 28.2, 28.4, 29.06, 29.14,
29.5, 31.7, 31.8, 33.5, 34.0, 39.3, 42.9, 52.5, 79.2, 80.3,
156.1, 180.6; IR (KBr) cmÀ1: 3211, 2979, 2926, 1726,
1687, 1519, 1461, 1388, 1249, 1178; HRMS calcd for
C15H29NO4Na (M+Na)+ 310.1994, found 310.2008.
Anal. calcd for C15H29NO4: C, 62.69; H, 10.17; N, 4.87;
Found: C 62.98; H, 10.51; N, 4.76; [a]2D5 +11.7 (c 1.64,
CHCl3).
4.1.7. Fragment B: Boc-L-Thr(Me)-OH (12b). Boc-l-
allo-Thr-OBn (2.19 g, 7.08 mmol) was dissolved in
CH2Cl2 (40 mL) and cooled at 0 ꢀC. To this solution
were added Me3OBF4 (1.57 g,10.6 mmol) and then
Proton-Sponge (2.28 g, 10.6 mmol). After being stirred
at 0 ꢀC for 2 h and at room temperature for an addi-
tional 10 h, the reaction mixture was filtered and the
filtrate was washed with 10% KHSO4, saturated
NaHCO3 aqueous solution and brine. The combined
organic layer was dried over Na2SO4, filtered and eva-
porated. The oil residue was purified by column chro-
matography with a 6:1 mixture of hexane and EtOAc as
an eluent to give benzyl ester of 12b (1.23 g, 3.80 mmol,
54%) as a colorless oil.
This ester (1.06 g, 3.28 mmol) was dissolved in CH3OH
(15 mL). To this solution, 10% Pd/C (100 mg) was
added and the resulting mixture was stirred at room
temperature for 4 h under H2 atmosphere. Pd/C was
removed by filtration and the filtrate was evaporated to
4.1.6. (R)-3-Hydroxynonanoic acid (11). Methyl 3-oxo-
nonanoate (10) (1.38 g, 7.41 mmol) was placed in a dry
Schlenk and dissolved in CH3OH (15 mL). To this
solution was added the in situ prepared (R)-BINAP-Ru
complex (24.5 mg) under N2 atmosphere. The resulting
solution was transferred by cannula to autoclave with
N2 gas. Hydrogen was introduced into the reaction ves-
sel until the pressure gauge indicated 6 atm. This mix-
ture was vigorously stirred at 100 ꢀC for 4 h. After the
reaction mixture was cooled, hydrogen was carefully
released. The mixture was evaporated and the oil resi-
due was purified by silica gel flash column chromato-
1
give 12b (0.76 g, 3.26 mmol, 99%) as a colorless oil. H
NMR (400 MHz, CD3OD) d ppm: 1.16 (d, 3H, J=6.3
Hz), 1.45 (s, 9H), 3.35 (s, 3H), 3.63–3.72 (m, 1H), 4.34
(d, 1H, J=4.7 Hz); 13C NMR (125 MHz, CD3OD) d
ppm: 15.8, 28.7, 57.0, 57.9, 78.2, 80.7, 97.3, 158.0, 174.0;
IR (CHCl3) cmÀ1: 3442, 2982, 2934, 1759, 1713, 1501,
1393, 1369, 1252, 1163; HRMS calcd for C10H20NO5
(M+H)+ 234.1341, found 234.1333; [a]2D4À3.9 (c 1.11,
CH3OH).

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
345
4.1.8. Boc-L-Thr(TBS)-Gly-OBn (14b). To the solution
of Boc-l-Threonine (12a) (2.00 g, 9.13 mmol) in CH2Cl2
(18 mL) were added Et3N (3.8 mL, 27.2 mmol) and
TBSOTf (5.2 mL, 22.6 mmol) at 0 ꢀC. The mixture was
stirred at the same temperature for 2 h. After being
stirred at room temperature for an additional 2 h, aqu-
eous K2CO3 solution was added. The resulting mixture
was vigorously stirred over night. The solution was
acidified with 10% aqueous KHSO4 solution and
extracted with CH2Cl2. The combined organic layer was
dried over Na2SO4, filtered and evaporated. The crude
oil was purified by column chromatography with a 4:1
mixture of hexane and EtOAc as an eluent to give Boc-
l-allo-Thr(TBS)-OH (2.69 g, 8.05 mmol, 88%) as a col-
orless solid (mp129–130 ꢀC).
4.1.10. Boc-L-Abu-Gly-OBn (14d). 97%, a colorless
crystal (mp, 80–81 ꢀC), H NMR (400 MHz, CDCl3) d
1
ppm: 0.95 (t, 3H, J=7.5 Hz), 1.44 (s, 9H), 1.59–1.71 (m,
1H), 1.83–1.94 (m, 1H), 4.03–4.15 (m, 1H), 4.09 (d, 2H,
J=5.5 Hz), 5.02 (br d, 1H, J=4.6 Hz), 5.18 (s, 2H), 6.64
(br s, 1H), 7.34–7.39 (m, 5H); 13C NMR (125 MHz,
CDCl3) d ppm: 9.9, 25.7, 28.3, 41.3, 55.7, 67.2, 80.1,
128.4, 128.55, 128.64, 135.1, 155.7, 169.5, 172.3; IR
(KBr) cmÀ1: 3321, 2970, 1739, 1684, 1658, 1555, 1529,
1278, 1254, 1172. Anal. calcd for C18H26N2O5: C, 61.70;
H, 7.48; N, 7.99; Found: C, 61.73; H, 7.57; N, 7.87;
HRMS calcd for C18H27N2O5 (M+H)+ 351.1920,
found 351.1912; [a]2D4 À20.2 (c 1.39, CHCl3).
4.1.11. Boc-L-allo-Thr(ꢀMe)-Gly-OBn (14e). 98%, a col-
orless solid (mp74–76 C), 1H NMR (400 MHz, CDCl3)
d ppm: 1.20 (d, 3H, J=6.3 Hz), 1.45 (s, 9H), 3.34 (s,
3H), 3.60–3.66 (m, 1H), 4.10 (d, 2H, J=5.2 Hz), 4.25
(br t, 1H, J=6.6 Hz), 5.15 (br s, 1H), 5.18 (s, 2H), 6.85
(br s, 1H), 7.30–7.41 (m, 5H); 13C NMR (125 MHz,
CDCl3) d ppm: 15.2, 28.2, 41.2, 41.3, 56.6, 67.2, 80.3,
128.4, 128.5, 128.6, 135.1, 155.8, 169.4, 170.6; IR (KBr)
cmÀ1: 3329, 2982, 2934, 1740, 1681, 1657, 1534, 1369,
1307, 1175; HRMS calcd for C19H29N2O6 (M+H)+
381.2026, found 381.2027. Anal. calcd for C19H28N2O6:
C, 59.99; H, 7.42; N, 7.36; Found: C 59.89; H, 7.42; N,
7.34; [a]2D4À7.4 (c1.27, CHCl3).
This product (2.46 g, 7.37 mmol) and glycine benzyl
ester p-toluenesulfonate (13) (2.50 g, 7.41 mmol) were
dissolved in CH2Cl2 (30 mL) and then Et3N (2.55 mL,
18.2 mmol) and 1-hydroxybenzotriazole (1.23 g, 9.10
mmol) were added. To the solution was added
ꢀ
.
EDCI HCl (1.55 g, 8.09 mmol) at 0 C. The reaction
mixture was stirred at 0 ꢀC for 2 h and at room tem-
perature for an additional 13 h. After a saturated
NaHCO3 aqueous solution was added, the organic layer
was separated and the aqueous layer was extracted with
CH2Cl2. The combined organic extracts were dried over
Na2SO4, filtered and evaporated. The residue was puri-
fied by column chromatography with a gradient elution
system, a 5:1–4:1 mixture of hexane and EtOAc as elu-
ents to give 14b (3.08 g, 6.41 mmol, 87%) as a colorless
4.1.12. Boc-L-Thr(TBS)-Gly-OH (15b). Compound 14b
(1.93 g, 4.02 mmol) was dissolved in C2H5OH (15 mL).
To this solution, 10% Pd/C (210 mg) was added and the
resulting mixture was stirred at room temperature for
2.5 h under H2 atmosphere. Pd/C was removed by fil-
tration and the filtrate was evaporated to give 15b (1.54
1
oil. H NMR (400 MHz, CDCl3) d ppm: 0.08 (s, 3H),
0.10 (s, 3H), 0.87 (s, 9H), 1.13 (d, 3H, J=6.2 Hz), 1.46
(s, 9H), 4.03 (dd, 1H, J=4.5, 18.5 Hz), 4.12–4.18 (m,
2H), 4.40 (dq, 1H, J=2.8, 6.3 Hz), 5.19 (s, 2H), 5.31 (d,
1H, J=6.3 Hz), 7.16 (br s, 1H), 7.31–7.39 (m, 5H); 13C
NMR (125 MHz, CDCl3) d ppm: À5.1, À4.7, 17.9, 18.9,
25.7, 28.3, 41.4, 59.4, 67.2, 68.3, 80.1, 128.4, 128.5,
1
g, 3.95 mmol, 98%) as a colorless amorphous foam. H
NMR (400 MHz, CD3OD) d ppm: 0.04 (s, 3H), 0.08 (s,
3H), 0.88 (s, 9H), 1.18 (d, 3H, J=6.3 Hz), 1.47 (s, 9H),
3.84 (d, 1H, J=17.9 Hz), 4.01 (d, 1H, J=17.9 Hz), 4.07
(d, 1H, J=3.0 Hz), 4.34 (dq, 1H, J=3.0, 6.3 Hz); 13C
NMR (125 MHz, CD3OD) d ppm: À4.9, À4.5, 18.8,
20.9, 26.3, 28.7, 41.9, 61.5, 70.2, 81.1, 157.8, 172.4,
173.2; IR (KBr) cmÀ1: 3332, 2955, 2931, 2858, 1722,
1668, 1505, 1255, 1170, 837; HRMS calcd for
C17H35N2O6Si (M+H)+ calcd 391.2264, found 391.2266.
128.6, 135.2, 155.8, 169.3, 170.4; IR (CHCl3) cmÀ1
:
3423, 2980, 2956, 2931, 2858, 1747, 1713, 1678, 1485,
1167; HRMS calcd for C24H41N2O6Si (M+H)+ calcd
481.2734, found 481.2735; [a]2D5 +3.5 (c 1.07, CHCl3).
4.1.9. Boc-L-allo-Thr(TBS)-N-Me-Gly-OBn (14c). 85%,
1
.
a slightly yellow oil, H NMR (400 MHz, CDCl3, two
Anal. calcd for C17H34N2O6Si 1/5H2O: C, 51.80; H,
8.80; N, 7.11; Found: C, 51.82; H, 8.52; N, 7.33;
rotamers (major/minor=2.7/1)) d ppm: À0.04* (s, 3/
4H), À0.03 (s, 9/4H), 0.03* (s, 3/4H), 0.04 (s, 9/4H),
0.83* (s, 9/4H), 0.84 (s, 27/4H), 1.21 (d, 3H, J=6.1 Hz),
1.40* (s, 9/4H), 1.42 (s, 27/4H), 2.98* (s, 3/4H), 3.19 (s,
9/4H), 3.75 (d, 3/4H, J=17.3 Hz), 3.81–3.90 (m, 5/4H),
4.39* (t, 1/4H, J=9.2 Hz), 4.58 (d, 3/4H, J=17.3 Hz),
6.25 (t, 3/4H, J=8.6 Hz), 4.73* (d, 1/4H, J=18.5 Hz),
5.15 and 5.16 (AB type d’s, each 1H, J=12.3 Hz), 5.25
(d, 1H, J=9.8 Hz), 7.30–7.39 (m, 5H); 13C NMR
(125 MHz, CDCl3, both rotamer) d ppm: À5.2, À5.0,
À4.81, À4.77, 14.1, 17.81, 17.84, 20.5, 20.7, 25.63, 25.67,
28.27, 28.31, 31.6, 35.2, 36.9, 49.6, 51.9, 55.4, 55.6, 66.9,
67.3, 70.8, 71.4, 77.5, 79.5, 79.6, 128.3, 128.4, 128.6,
135.2, 135.3, 155.0, 155.2, 168.61, 168.64, 172.5, 172.8;
IR (CHCl3) cmÀ1: 3434, 2957, 2931, 2858, 1749, 1710,
1650, 1499, 1258, 1170; HRMS calcd for C25H43N2O6Si
(M+H)+ calcd 495.2891, found 495.2874; [a]2D5 +13.1
(c 1.12, CHCl3).
[a]2D4=À0.7 (c 1.09, CH3OH).
4.1.13. Boc-L-allo-Thr(TBS)-N-Me-Gly-OH (15c). 99%,
a colorless amorphous foam, 1H NMR (400 MHz,
CD3OD, two rotamers (major/minor=2.0/1)) d ppm:
0.004 (s, 3H), 0.07 (s, 3H), 0.87 (s, 9H), 1.17–1.20 (m,
3H), 1.41* (s, 3H), 1.43 (s, 6H), 2.96* (s, 1H), 3.23 (s,
2H), 3.80 (d, 2/3H, J=17.2 Hz), 3.92–4.05 (m, 4/3H),
4.36 (d, 1H, J=17.2 Hz), 4.53–4.61 (m, 1H); 13C NMR
(125 MHz, CD3OD, both rotamer) d ppm: À4.8, À4.5,
À4.4, À3.6, 14.4, 18.7, 20.9, 23.7, 26.2, 26.3, 28.0, 28.7,
32.7, 35.6, 37.5, 50.7, 52.5, 57.1, 70.9, 71.5, 80.6, 80.7,
97.3, 157.5, 172.0, 172.1, 174.3, 174.4; IR (CHCl3)
cmÀ1: 3432, 2981, 2957, 2931, 2859, 1711, 1651, 1500,
1257, 1164; HRMS calcd for C18H37N2O6Si (M+H)+
calcd 405.2421, found 405.2447. Anal. calcd for
.
C18H36N2O6Si 1/5H2O: C, 52.97; H, 8.99; N, 6.86;

346
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
Found: C, 52.82; H, 9.07; N, 6.65; [a]2D5À2.1 (c 1.11,
(mp56–57 ꢀC). 1H NMR (400 MHz, CDCl3) d ppm:
0.84 (d, 3H, J=7.1 Hz), 0.92 (t, 3H, J=7.4 Hz), 1.44 (s,
9H), 1.92 (br s, 1H), 3.29 (s, 3H), 3.60 (dd, 1H, J=3.1,
9.4 Hz), 3.84 (dd, 1H, J=3.0, 9.4 Hz), 4.18 (brd, 1H,
J=3.8 Hz), 4.76–4.80 (m, 1H), 5.05 (br d, 1H, J=8.1
Hz), 5.14 and 5.26 (AB type d’s, each 1H, J=12.3 Hz),
6.64 (br d, 1H, J=6.9 Hz), 7.39–7.38 (m, 5H); 13C
NMR (125 MHz, CDCl3) d ppm: 11.7, 14.1, 26.3, 38.3,
37.8, 52.7, 57.9, 59.2, 67.3, 72.1, 79.9, 128.2, 128.4,
128.6, 135.3, 155.8, 169.9, 171.6; IR (KBr) cmÀ1: 3321,
2967, 2932, 1746, 1658, 1522, 1366, 1243, 1168; HRMS
calcd for C22H35N2O6 (M+H)+ 423.2495, found
423.2495. Anal. calcd for C22H34O6N2: C, 62.54; H,
8.11; N, 6.03; Found: C, 62.43; H, 7.89; N, 6.71; [a]D24
+3.5 (c 1.44, CHCl3).
CH3OH).
4.1.14. Boc-L-Abu-Gly-OH (15d). 100%, a colorless
amorphous foam, 1H NMR (400 MHz, CD3OD) d ppm:
0.97 (t, 3H, J=7.4 Hz), 1.44 (s, 9H), 1.56–1.68 (m, 1H),
1.77–1.87 (m, 1H), 3.86 and 3.96 (AB type d’s, each 1H,
J=17.8 Hz), 3.97–4.00 (m, 1H); 13C NMR (125 MHz,
CD3OD) d ppm: 10.6, 26.7, 28.7, 41.8, 57.4, 80.6, 157.9,
172.7, 175.5; IR (CHCl3) cmÀ1: 3433, 3368, 2980, 2936,
1730, 1672, 1502, 1370, 1251, 1162; HRMS calcd for
C11H21N2O5 (M+H)+ 261.1450, found 261.1463; [a]2D4
À26.4 (c 1.18, CH3OH).
4.1.15. Boc-L-allo-Thr(Me)-Gly-OH (15e). 100%, a col-
ꢀ
1
orless solid, (mp50 C), H NMR (400 MHz, CD3OD)
d ppm: 1.15 (d, 3H, J=6.3 Hz), 1.45 (s, 9H), 3.35 (s,
3H), 3.65–3.68 (m, 1H), 3.92 and 3.94 (AB type d’s,
each 1H, J=17.8 Hz), 4.33 (br d, 1H, J=4.8 Hz), 13C
NMR (125 MHz, CD3OD) d pm: 15.5, 28.7, 41.8, 57.0,
58.5, 78.1, 80.8, 97.3, 157.9, 172.6, 173.3; IR (KBr)
cmÀ1: 3322, 3091, 2980, 2935, 1719, 1663, 1528, 1368,
1248, 1168; HRMS calcd for C12H23N2O6 (M+H)+
291.1556, found 291.1565. Anal. calcd for C12H22N2O6:
C, 49.65; H, 7.64; N, 9.65; Found: C 49.36; H, 7.85; N,
9.43; [a]2D5À13.0 (c 1.03, CH3OH).
4.1.18. Boc-L-Ile-L-Ser(Bn)-OAllyl (18g). 99%, a color-
ꢀ
less solid, (mp59–60 C), 1H NMR (400 MHz, CDCl3) d
ppm: 0.91 (t, 3H, J=7.4 Hz), 0.95 (d, 3H, J=6.7 Hz),
1.10–1.21 (m, 1H), 1.40–1.52 (m, 1H), 1.45 (s, 9H),
1.85–1.92 (m, 1H), 3.68 (dd, 1H, J=3.3, 9.5 Hz), 3.93
(dd, 1H, J=3.0, 9.5 Hz), 4.04 (br t, 1H, J=6.5 Hz), 4.48
and 4.55 (AB type d’s, each 1H, J=12.2 Hz), 4.60–4.69
(m, 2H), 4.77 (dt, 1H, J=3.0, 8.0 Hz), 5.08 (br d, 1H,
J=7.9 Hz), 5.23 (dt, 1H, J=1.2, 10.3 Hz), 5.31 (dt, 1H,
J=1.3, 17.5 Hz), 5.82–9.92 (m, 1H), 6.63 (br d, 1H,
J=7.5 Hz), 7.26–7.36 (m, 5H); 13C NMR (125 MHz,
CDCl3) d ppm: 11.5, 15.4, 24.6, 28.3, 37.7, 52.6, 59.1,
66.2, 69.6, 73.3, 79.8, 118.8, 127.7, 127.9, 128.4, 131.5,
137.4, 155.6, 169.6, 171.2; IR (KBr) cmÀ1: 3334, 2966,
2934, 1747, 1687, 1652, 1548, 1524, 1245, 1173; HRMS
calcd for C24H37N2O6 (M+H)+ calcd 449.2652, found
449.2651. Anal. calcd for C24H36N2O6: C, 64.26; H,
8.09; N, 6.25; Found: C, 64.48; H, 8.15; N, 6.25; [a]D27
+8.6 (c 1.01, CHCl3).
.
4.1.16. Fragment C: L-Ser(Me)-OBn HCl (17a). The
benzyl ester (2.87 g, 9.28 mmol) prepared from Boc-l-
Ser(Me)-OH (16a) was treated with 4 N HCl in EtOAc
(40 mL) and the mixture was stirred at room tempera-
ture for 2 h. The solvents were removed in vacuo and
dissolved in EtOAc. The mixture was evaporated and
dissolved in EtOAc again. This procedure was repeated
tree times and the resulting crude product was dried
under reduced pressure to give a hydrochloride salt
(2.28 g, 9.28 mmol, 100%) as a colorless solid (mp101–
4.1.19. Boc-19a. Deprotection procedure of Boc group
in 18a was almost the same in the synthesis of 17a to
give a hydrochloride salt (3.00 g) as a colorless solid (mp
169 ꢀC).
102 ꢀC). H NMR (400 MHz, CDCl3) d ppm: 3.34 (s,
1
3H), 3.92 (dd, 1H, J=3.6, 10.4 Hz), 4.03 (dd, 1H,
J=2.7, 10.4 Hz), 4.45 (br s, 1H), 5.16 and 5.29 (AB type
d’s, each 1H, J=12.2 Hz), 7.28–7.37 (m, 5H), 8.77 (br s,
1H); 13C NMR (125 MHz, CDCl3) d ppm: 53.6, 59.3,
68.2, 69.2, 128.3, 128.58, 128.60, 134.7, 167.5; IR (KBr)
cmÀ1: 2941, 2883, 1963, 1753, 1576, 1487, 1225, 1107,
968; HRMS calcd for C11H16NO3 (M+H)+ 210.1130,
found 210.1119. Anal. calcd for C11H16ClNO3: C,
53.77; H, 6.56; N, 5.74; Cl, 14.43; Found: C, 53.44; H,
6.59; N, 5.74; Cl, 14.72; [a]2D5À23.1 (c 1.15, CHCl3).
This salt (1.53 g, 4.28 mmol) and Boc-N-Me-l-leucine
(1.10 g, 4.49 mmol) were dissolved in CH2Cl2 (15 mL).
To this solution were added Et3N (1.43 mL, 10.3 mmol)
and PyBOP (2.34 g, 4.49 mmol) at 0 ꢀC. The reaction
mixture was stirred at 0 ꢀC for 2 h and at room tem-
perature for an additional 10 h. After a saturated
NaHCO3 aqueous solution was added, the organic layer
was separated and the aqueous layer was extracted with
CH2Cl2. The combined organic extracts were dried over
Na2SO4, filtered and evaporated. The oil residue was
purified by column chromatography with a 2:1 mixture
of hexane and EtOAc as an eluent to give Boc-19a (2.26
g, 41.1 mmol, 96%) as a colorless oil. 1H NMR
(400 MHz, CDCl3) d ppm: 0.83 (d, 3H, J=6.8 Hz), 0.89
(t, 3H, J=6.9 Hz), 0.91 (d, 3H, J=6.5 Hz), 0.95 (d, 3H,
J=6.6 Hz), 1.06–1.17 (m, 1H), 1.33–1.46 (m, 2H), 1.48
(s, 9H), 1.65–1.71 (m, 2H), 1.83–1.97 (m, 1H), 2.75 (s,
3H), 3.29 (s, 3H), 3.58 (dd, 1H, J=3.2, 9.0 Hz), 3.84 (br
d, 1H, J=8.3 Hz), 4.49 (dd, 1H, J=4.4, 9.0 Hz), 4.63–
4.68 (m, 1H), 4.75–4.77 (m, 1H), 5.13 and 5.25 (AB type
d’s, each 1H, J=12.2 Hz), 6.47 (br s, 1H), 6.66–6.74 (m,
1H), 7.31–7.40 (m, 5H); 13C NMR (125 MHz, CDCl3)
4.1.17. Boc-L-allo-Ile-L-Ser(Me)-OBn (18a). Compound
17 (2.22 g, 9.04 mmol) and Boc-l-Ile-OH (1.99 g, 8.60
mmol) were dissolved in CH2Cl2 (30 mL). To this solu-
tion were added Et3N (3.02 mL, 21.7 mmol) and PyBOP
(4.70 g, 9.04 mmol) at 0 ꢀC. The reaction mixture was
stirred at 0 ꢀC for 2 h and at room temperature for an
additional 10 h. After saturated NaHCO3 aqueous
solution was added, the organic layer was separated and
the aqueous layer was extracted with CH2Cl2. The
combined organic extracts were dried over Na2SO4, fil-
tered and evaporated. The oil residue was purified by
column chromatography with a gradient elution system,
a 3:1–1:1 mixture of hexane and EtOAc as eluents to
give 18a (3.63 g, 8.61 mmol, 95%) as a colorless solid

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
347
d ppm: 11.7, 14.2, 22.1, 23.1, 23.2, 24.5, 24.8, 26.3, 28.4,
29.6, 29.9, 36.2, 36.3, 37.3, 52.6, 56.1, 59.2, 67.3, 72.0,
80.5, 128.2, 128.4, 128.6, 135.3, 156.8, 169.8, 171.0,
170.1, 171.5; IR (CHCl3) cmÀ1: 3432, 296ss4, 2934,
1746, 1676, 1501, 1390, 1323, 1152, 1118; HRMS calcd
for C29H48N3O3 (M+H)+ 550.3492, foundd 550.3462;
[a]2D6 À61.3 (c 1.27, CHCl3).
5.81–5.91 (m, 1H), 6.62 (d, 1H, J=8.2 Hz), 7.25–7.36
(m, 5H), 7.88 (d, 1H, J=9.1 Hz); 13C NMR (125 MHz,
CDCl3) d ppm: 11.7, 14.3, 15.4, 21.8, 23.2, 25.2, 26.4,
37.6, 43.1, 43.2, 52.6, 55.8, 61.6, 66.2, 69.6, 73.3, 118.7,
127.6, 127.8, 128.4, 131.5, 137.4, 169.6, 171.2; IR (KBr)
cmÀ1: 3282, 3070, 2959, 2872, 1740, 1633, 1551, 1393,
1215, 1102; HRMS calcd for C27H44N3O5 (M+H)+
490.3281, found 490.3291. Anal. calcd for C27H43N3O5:
C, 66.23; H, 8.85; N, 8.58; Found: C 66.03; H, 8.64; N,
8.68; [a]2D5 À11.7 (c 1.04, CHCl3).
4.1.20. N-Me-L-Leu-L-allo-Ile-L-Ser(Me)-OBn (19a).
Compound Boc-19a (2.19 g, 3.98 mmol) was treated
with 4N HCl in EtOAc (20 mL) and the mixture was
stirred at room temperature for 1.3 h. The reaction
mixture was concentrated under reduced pressure, dilu-
ted with EtOAc and washed with saturated NaHCO3
aqueous solution. The organic layer was dried over
Na2SO4, filtered and evaporated to give 19a (1.66 mg,
3.69 mmol, 93%) as a colorless solid (mp, 110–111 ꢀC).
This free amine was directly used in the following reac-
4.1.23. Boc-19d. 95%, a colorless solid (mp, 108–
ꢀ
1
109 C), H NMR (400 MHz, CDCl3) d ppm: 0.90–0.94
(m, 12H), 1.12–1.23 (m, 1H), 1.38–1.53 (m, 2H), 1.44 (s,
9H), 1.63–1.68 (m, 2H), 1.96 (br, 1H), 3.65 (dd, 1H,
J=3.2, 9.5 Hz), 3.93 (dd, 1H, J=3.1, 9.5 Hz), 4.11 (br,
1H), 4.49 (dd, 1H, J=4.8, 8.9 Hz), 4.47 and 4.56 (AB
type d’s, each 1H, J=12.1 Hz), 4.59–4.68 (m, 2H), 4.75
(dt, 1H, J=3.4, 8.4 Hz), 4.88 (br d, 1H, J=7.3 Hz), 5.23
(dd, 1H, J=1.3, 10.6 Hz), 5.30 (dt, 1H, J=1.3, 16.9
Hz), 5.81–5.91 (m, 1H), 6.63 (br d, 1H, J=7.6 Hz), 6.68
(br d, 1H, J=8.2 Hz), 7.26–7.37 (m, 5H); 13C NMR
(125 MHz, CDCl3) d ppm: 11.7, 14.2, 22.0, 22.9, 24.7,
26.1, 28.3, 37.8, 40.6, 52.6, 53.2, 56.4, 66.2, 69.5, 73.3,
80.1, 118.8, 127.7, 127.9, 128.4, 131.5, 137.3, 155.7,
169.5, 171.8, 172.5; IR (KBr) cmÀ1: 3283, 2961, 2932,
2874, 1745, 1695, 1644, 1543, 1523, 1173; HRMS calcd
for C30H48N3O7 (M+H)+ calcd 562.3492, found
562.3492. Anal. calcd for C30H47N3O7: C, 64.15; H,
8.43; N, 7.48; Found: C, 64.11; H, 8.65; N, 7.53;
[a]2D8=À23.5 (c 1.18, CHCl3).
1
tion without further purification. H NMR (400 MHz,
CD3OD) d ppm: 0.86–0.91 (m, 12H), 0.94 (d, 1H,
J=6.6 Hz), 1.10–1.21 (m, 1H), 1.35–1.52 (m, 3H), 1.60–
1.70 (m, 1H), 1.84–1.94 (m, 1H), 2.31 (s, 3H), 3.13 (t,
1H, J=7.2 Hz), 3.32 (s, 3H), 3.62 (dd, 1H, J=3.7, 9.6
Hz), 3.80 (dd, 1H, J=5.0, 9.6 Hz), 4.51 (d, 1H, J=6.0
Hz), 4.67 (t, 1H, J=4.3 Hz), 4.13 and 5.21 (AB type d’s,
each 1H, d, J=12.2 Hz), 7.29–7.40 (m, 5H); 13C NMR
(125 MHz, CD3OD) d ppm: 11.9, 14.8, 22.9, 23.1, 25.7,
27.4, 33.7, 38.6, 42.8, 54.2, 57.9, 59.3, 63.0, 68.1, 72.7,
129.28, 129.33, 129.5, 137.1, 171.1, 173.3; IR (KBr)
cmÀ1: 3287, 2960, 2930, 1742, 1634, 1547, 1456, 1390,
1208, 1118; HRMS calcd for C24H40N3O5 (M+H)+
450.5916, found 450.2954. Anal. calcd for C24H39N3O5:
C, 64.12; H, 8.74; N, 9.35; Found: C, 64.44; H, 8.72; N,
9.36; [a]2D5 À23.0 (c 1.07, CHCl3).
4.1.21. Boc-19c. 94%, a colorless oil, 1H NMR
(400 MHz, CD3OD) d ppm: 0.88–0.95 (m, 12H), 1.11 (t,
3H, J=7.0 Hz), 1.24–1.34 (m, 2H), 1.45–1.55 (m, 2H),
1.49 (s, 9H), 1.64–1.67 (m, 1H), 1.88–1.98 (m, 1H),
3.15–3.30 (m, 2H), 3.74 (dd, 1H, J=3.7, 9.6 Hz), 3.89
(dd, 1H, J=5.1, 9.6 Hz), 4.47 (d, 1H, J=5.1 Hz), 4.48
and 4.57 (AB type d’s, each 1H, J=11.8 Hz), 4.62–4.64
(m, 3H), 4.69 (dd, 1H, J=3.8, 4.8 Hz), 5.20 (dd, 1H,
J=1.4, 11.6 Hz), 5.32 (dd, 1H, J=1.4, 17.6 Hz), 5.85–
5.95 (m, 1H), 7.27–7.33 (m, 5H); 13C NMR (125 MHz,
CD3OD) d ppm: 12.0, 14.4, 14.8, 23.3, 23.7, 26.0, 27.4
28.7, 32.7, 38.7, 54.3, 57.6, 67.0, 70.4, 74.2, 97.3, 118.7,
128.8, 128.9, 129.4, 133.2, 139.1, 170.9, 173.6; IR
(CHCl3) cmÀ1: 3432, 2964, 2874, 1746, 1676, 1502, 1368,
1287, 1156, 1115; HRMS calcd for C32H51N3O7Na
(M+Na)+ 612.3625, found 612.3635; [a]2D4 À49.4 (c
1.38, CHCl3).
4.1.24. L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (19d). 98%, a
colorless solid (mp, 90–91 ꢀC), 1H NMR (400 MHz,
CDCl3) d ppm: 0.89–0.99 (m, 12H), 1.13–1.24 (m, 1H),
1.31–1.39 (m, 1H), 1.41–1.51 (m, 1H), 1.57–1.80 (m,
4H), 1.93–2.02 (m, 1H), 3.43 (dd, 1H, J=3.1, 9.5 Hz),
3.68 (dd, 1H, J=3.1, 9.5 Hz), 3.93 (dd, 1H, J=3.1, 9.5
Hz), 4.46 (dd, 1H, J=5.1, 9.1 Hz), 4.48 and 4.55 (AB
type d’s, each 1H, J=12.2 Hz), 4.59–4.69 (m, 2H), 4.75
(dt, 1H, J=3.1, 8.0 Hz), 5.24 (dd, 1H, J=1.2, 10.3 Hz),
5.29–5.34 (m, 1H), 5.82–5.92 (m, 1H), 6.69 (d, 1H,
J=8.1 Hz), 7.26–7.36 (m, 5H), 7.86 (d, 1H, J=9.1 Hz);
13C NMR (125 MHz, CDCl3) d ppm: 11.7, 14.4, 21.3,
23.4, 24.9, 26.3, 37.6, 44.1, 52.6, 53.7, 56.1, 66.2, 69.5,
73.3, 118.8, 127.7, 127.9, 128.4, 131.4, 137.3, 169.7,
171.3, 175.7; IR (KBr) cmÀ1: 3299, 2959, 2932, 2873,
1743, 1636, 1546, 1237, 1210, 1106; HRMS calcd for
C25H40N3O5 (M+H)+ calcd 462.2968, found 462.2975.
Anal. calcd for C25H39N3O5: C, 65.05; H, 8.52; N, 9.10;
Found: C, 64.84; H, 8.65; N, 8.90; [a]2D3 –13.3 (c 1.08,
CHCl3).
4.1.22. N-Et-L-Leu-L-allo-Ile-L-Ser(Me)-OBn (19c). 97%,
a colorless solid (mp112–113 ꢀC), H NMR (400 MHz,
4.1.25. Boc-19e. 97%, a colorless solid, (mp56–58 ꢀC),
1H NMR (400 MHz, CDCl3) d ppm: 0.87–0.91 (m, 6H),
1.11–1.22 (m, 1H), 1.47 (s, 9H), 1.76 (br s, 1H), 1.84–
2.20 (br m, 4H), 2.37 (br s, 1H), 3.31–3.55 (br m, 2H),
3.69 (br d, 1H, J=6.8 Hz), 3.91 (br d, 1H, J=7.2 Hz),
4.25 (br s, 1/2H), 4.35 (br s, 1/2H), 4.44–4.52 (m, 2H),
4.56 (d, 1H, J=12.2 Hz), 4.59–4.68 (m, 2H), 4.76 (dt,
1H, J=3.2, 8.1 Hz), 5.23 (d, 1H, J=10.2 Hz), 5.28–5.33
(m, 1H), 5.82–5.91 (m, 1H), 6.50 (br s, 1/4H), 6.64 (br s,
1/4H), 6.84 (br s, 1/2H), 7.26–7.36 (m, 11/2H), 7.47 (br
1
CDCl3) d ppm: 0.90 (d, 3H, J=6.7 Hz), 0.92–1.01 (m,
9H), 1.08 (t, 3H, J=7.0 Hz), 1.11–1.26 (m, 1H), 1.35–
1.49 (m, 2H), 1.56–1.77 (m, 3H), 1.94–2.06 (m, 1H),
2.55–2.68 (m, 2H), 3.14 (dd, 1H, J=4.5, 9.2 Hz), 3.67
(dd, 1H, J=3.2, 9.3 Hz), 3.93 (dd, 1H, J=3.2, 9.3 Hz),
4.46–4.49 (m, 1H), 4.47 and 4.56 (AB type d’s, each 1H,
J=12.2 Hz), 4.59–4.73 (m, 2H), 4.75–4.79 (m, 1H), 5.23
(dd, 1H, J=1.3, 8.5 Hz), 5.30 (dd, 1H, J=1.3, 17.5 Hz),

348
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
s, 1/2H); 13C NMR (125 MHz, CDCl3) d ppm: 11.7,
14.1, 24.7, 26.3, 27.8, 28.4, 37.0, 47.0, 47.1, 52.6, 56.7,
59.9, 66.1, 69.6, 73.3, 80.5, 118.7, 127.7, 127.8, 128.4,
118.8, 127.7, 127.9, 128.5, 131.5, 137.4, 169.7, 171.2,
174.2; IR (KBr) cmÀ1: 3278, 2959, 2922, 2875, 2853,
1738, 1636, 1544, 1452, 1220; HRMS calcd for
C25H38N3O5 (M+H)+ calcd 460.2812, found 460.2783.
Anal. calcd for C25H37N3O5: C, 65.34; H, 8.12; N, 9.14;
Found: C, 65.62; H, 8.02; N, 9.10; [a]2D6 À13.4 (c 1.05,
CHCl3).
131.5, 137.5, 169.6, 171.1, 172.1; IR (CHCl3) cmÀ1
:
2979, 2936, 1744, 1679, 1514, 1503, 1460, 1394, 1271,
1168; HRMS calcd for C29H44N3O7 (M+H)+ calcd
546.3179, found 546.3181; [a]2D4 À51.3 (c 1.06, CHCl3).
4.1.26. L-Pro-L-allo-Ile-L-Ser(Bn)-OAllyl (19e). 95%, a
colorless solid, (mp, 93–95 ꢀC), 1H NMR (400 MHz,
CDCl3) d ppm: 0.89 (d, 3H, J=6.8 Hz), 0.93 (t, 3H,
J=7.4 Hz), 1.11–1.22 (m, 1H), 1.39–1.49 (m, 1H), 1.71
(qu, 2H, J=6.8 Hz), 1.88–2.19 (m, 4H), 2.92 (dt, 1H,
J=6.2, 10.2 Hz), 3.03 (dt, 1H, J=6.9, 10.2 Hz), 3.68
(dd, 1H, J=3.4, 9.5 Hz), 3.76 (dd, 1H, J=5.0, 9.2 Hz),
3.93 (dd, 1H, J=3.4, 9.5 Hz), 4.40 (dd, 1H, J=5.1, 9.1
Hz), 4.47 and 4.55 (AB type d’s, each 1H, J=12.2 Hz),
4.59–4.69 (m, 2H), 4.75 (dt, 1H, J=3.4, 8.0 Hz), 5.27
(dd, 1H, J=1.4, 8.5 Hz), 5.29–5.34 (m, 1H), 5.82–5.92
(m, 1H), 6.71 (d, 1H, J=8.1 Hz), 7.26–7.37 (m, 5H),
8.19 (d, 1H, J=9.3 Hz); 13C NMR (125 MHz, CDCl3) d
ppm: 11.7, 14.4, 26.2, 26.4, 31.0, 37.5, 47.3, 52.7, 56.1,
60.6, 66.2, 69.6, 73.3, 118.8, 127.7, 127.9, 128.4, 131.5,
137.4, 169.7, 171.3, 175.3; IR (CHCl3) cmÀ1: 3433, 3326,
2968, 2936, 2876, 1746, 1665, 1511, 1502, 1105; HRMS
calcd for C24H36N3O5 (M+H)+ calcd 446.2655, found
446.2653. Anal. calcd for C24H35N3O5: C, 64.70; H,
7.92; N, 9.43; Found: C, 64.39; H, 7.75; N, 9.32; [a]D27
À24.3 (c 1.01, CHCl3).
4.1.27. Boc-19f. 97%, a colorless oil, 1H NMR
(400 MHz, CDCl3) d ppm: 0.88 (d, 3H, J=6.8 Hz), 0.94
(t, 3H, J=7.3 Hz), 1.14–1.21 (m, 1H), 1.42–1.56 (m,
4H), 1.48 (s, 9H), 1.60–1.72 (br, 2H), 1.95 (br, 1H), 2.27
(br, 1H), 2.80 (br, 1H), 3.65 (dd, 1H, J=3.0, 9.5 Hz),
3.93 (dd, 1H, J=3.0, 9.5 Hz), 4.02 (br s, 1/2H), 4.14 (br
s, 1/2H), 4.49 (br s, 1H), 4.47 and 4.56 (AB type d’s,
each 1H, J=12.2 Hz), 4.58–4.68 (m, 2H), 4.76 (dt, 2H,
J=3.0, 8.2 Hz), 5.23 (dt, 1H, J=1.3, 10.3 Hz), 5.31 (dq,
1H, J=1.4, 16.9 Hz), 5.81–5.91 (m, 1H), 6.60 (br s, 3/
2H), 6.79 (br s, 1/2H), 7.26–7.37 (m, 5H); 13C NMR
(125 MHz, CDCl3) d ppm: 11.8, 14.1, 14.5, 20.5, 22.7,
25.0, 26.4, 28.4, 31.6, 52.5, 52.6, 56.4, 66.2, 69.5, 73.4,
77.5, 80.8, 118.8, 127.7, 127.9, 128.5, 131.5, 137.3, 169.6,
170.8; IR (CHCl3) cmÀ1: 3432, 2968, 2939, 2876, 1746,
1676, 1501, 1411, 1367, 1161; HRMS calcd for
C30H46N3O7 (M+H)+ calcd 560.3335; Found
560.3340; [a]2D4 À48.6 (c 1.13, CHCl3).
4.1.29. Boc-19g. 100%, a colorless oil, 1H NMR
(400 MHz, CDCl3) d ppm: 0.89–0.93 (m, 12H), 1.12–
1.20 (m, 1H), 1.43–1.55 (m, 2H), 1.49 (s, 9H), 1.60–1.75
(m, 2H), 1.82–1.94 (m, 1H), 2.76 (s, 3H), 3.66 (dd, 1H,
J=3.4, 9.5 Hz), 3.93 (br d, 1H, J=8.2 Hz), 4.35 (t, 1H,
J=6.5 Hz), 4.47 and 4.56 (AB type d’s, each 1H,
J=12.2 Hz), 4.59–4.72 (m, 3H), 4.74–4.77 (m, 1H), 5.23
(d, 1H, J=10.1 Hz), 5.30 (dt, 1H, J=1.3, 15.6 Hz),
5.81–5.91 (m, 1H), 6.49 (br s, 1H), 6.62–6.73 (br m, 1H),
7.26–7.37 (m, 5H); 13C NMR (125 MHz, CDCl3) d ppm:
11.3, 15.4, 21.9, 23.1, 24.5, 24.8, 28.3, 29.9, 36.1, 37.1,
52.6, 57.4, 66.2, 69.5, 73.3, 80.4, 118.8, 127.7, 127.9,
128.4, 131.5, 137.3, 169.5, 170.6, 171.3, 171.5; IR
(CHCl3) cmÀ1: 3432, 2964, 2934, 2874, 1746, 1676, 1501,
1391, 1368, 1153; HRMS calcd for C31H50N3O7
(M+H)+ calcd 576.3649, found 576.3638; [a]2D4 À56.7 (c
1.08, CHCl3).
4.1.30. N-Me-L-Leu-L-Ile-L-Ser(Bn)-OAllyl (19g). 94%,
a colorless solid, (mp91–92 ꢀC), H NMR (400 MHz,
1
CDCl3) d ppm: 0.89–0.97 (m, 12H), 1.11–1.22 (m, 1H),
1.35–1.43 (m, 1H), 1.46–1.61 (m, 3H), 1.64–1.74 (m,
1H), 1.89–1.99 (m, 1H), 2.36 (s, 3H), 3.00 (dd, 1H,
J=4.9, 9.2 Hz), 3.66 (dd, 1H, J=3.1, 9.5 Hz), 3.93 (dd,
1H, J=3.0, 9.5 Hz), 4.35 (dd, 1H, J=6.4, 9.2 Hz), 4.47
and 4.55 (AB type d’s, each 1H, J=12.2 Hz), 4.59–4.68
(m, 2H), 4.76 (dt, 1H, J=3.3, 8.2 Hz), 5.23 (dt, 1H,
J=1.3, 9.9 Hz), 5.30 (dt, 1H, J=1.3, 17.5 Hz), 5.81–
5.91 (m, 1H), 6.64 (d, 1H, J=8.2 Hz), 7.26–7.36 (m,
5H), 7.70 (d, 1H, J=9.2 Hz); 13C NMR (100 MHz,
CDCl3) d ppm: 11.3, 15.6, 21.9, 23.2, 24.7, 25.2, 35.5,
37.3, 43.0, 52.6, 57.0, 63.7, 66.2, 69.6, 73.4, 118.8, 127.7,
127.9, 128.5, 131.5, 137.4, 169.6, 171.1, 174.9; IR
(CHCl3) cmÀ1: 3290, 2961, 2933, 2874, 1744, 1651, 1633,
1547, 1211, 1114; HRMS calcd for C26H42N3O5
(M+H)+ calcd 476.3125, found 476.3130. Anal. calcd
.
for C26H41N3O5 1/5H2O: C, 65.16; H, 8.71; N, 8.77;
Found: C, 65.23; H, 8.51; N, 8.90; [a]2D4 À21.3 (c 1.06,
CHCl3).
4.1.31. (2R,3R)-3-Hydroxy-2-methyltridecanoyl-N-Me-L-
Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (20c). Compound 4c
(179 mg, 0.732 mmol) and 19b (289 mg, 0.608 mmol)
were dissolved in THF (4.0 mL) and then Et3N (0.17
mL, 1.22 mmol) was added. To the solution was added
DEPC (149 mL, 0.913 mmol) at 0 ꢀC. The reaction mix-
ture was stirred at 0 ꢀC for 2 h and at room temperature
for additional 14 h. After saturated NaHCO3 aqueous
solution was added, the organic layer was separated and
the aqueous layer was extracted with EtOAc. The com-
bined organic extracts were dried over Na2SO4, filtered
and evaporated. The oil residue was purified by column
chromatography with a gradient elution system, a 4:1–
2:1 mixture of hexane and EtOAc used as eluents to give
4.1.28. L-homo-Pro-L-allo-Ile-L-Ser(Bn)-OAllyl (19f).
97%, a colorless solid, (mp111–112 ꢀC), 1H NMR
(400 MHz, CDCl3) d ppm: 0.92 (d, 3H, J=6.8 Hz), 0.94
(t, 3H, J=7.4 Hz), 1.16–1.26 (m, 1H), 1.37-1.59 (m,
5H), 1.75–1.81 (m, 1H), 1.91–2.05 (m, 3H), 2.67 (dt, 1H,
J=3.1, 11.7 Hz), 3.02 (dt, 1H, J=2.9, 11.7 Hz), 3.25
(dd, 1H, J=3.1, 9.6 Hz), 3.67 (dd, 1H, J=3.3, 9.5 Hz),
3.93 (dd, 1H, J=3.4, 9.5 Hz), 4.48 (dd, 1H, J=5.3, 9.0
Hz), 4.47 and 4.55 (AB type d’s, each 1H, J=12.0 Hz),
4.59–4.69 (m, 2H), 4.74 (dt, 1H, J=3.1, 8.1 Hz), 5.24
(dt, 1H, J=1.3, 11.5 Hz), 5.29–5.34 (m, 1H), 5.82–5.92
(m, 1H), 6.65 (d, 1H, J=8.1 Hz), 7.23–7.37 (m, 6H); 13C
NMR (125 MHz, CDCl3) d ppm: 11.7, 14.5, 23.9, 25.8,
26.3, 30.1, 37.7, 45.6, 52.6, 56.0, 60.2, 66.2, 69.5, 73.3,
1
20c (425 mg, 0.605 mmol, 99%) as a colorless oil. H

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
349
NMR [400 MHz, CDCl3, two rotamers (major/
minor=2/1)] d ppm: 0.80–0.97 (m, 15H), 1.07* (d, 1H,
J=6.6 Hz), 1.10-1.20 (m, 1H), 1.22–1.40 (m, 18H),
1.41–1.53 (m, 3H), 1.58–1.75 (m, 1H), 1.77–1.85 (m, 5/
3H), 1.94–2.00 (m, 4/3H), 2.69–2.77 (m, 2/3H), 2.80* (s,
1H), 2.84–2.91* (m, 1/3H), 2.95 (s, 2H), 3.25* (br s, 1/
3H), 3.60–3.64 (m, 2/3H), 3.68 (dd, 2/3H, J=3.3, 9.5
Hz), 3.69* (dd, 1/3H, J=3.1, 9.4 Hz), 3.70–3.77 (br,
1H), 3.93 (dd, 1H, J=3.0, 9.5 Hz), 4.44–4.57 (m, 4/3H),
4.48 and 4.55 (AB type d’s, each 1H, J=12.2 Hz), 4.59–
4.68 (m, 2H), 4.73–4.78 (m, 1H), 5.10 (dd, 2/3H, J=5.7,
10.1 Hz), 5.24 (dd, 1H, J=1.2, 8.6 Hz), 5.31 (dd, 1H,
J=1.3, 14.1 Hz), 5.82–5.92 (m, 1H), 6.64 (d, 2/3H,
J=8.2 Hz), 6.80–6.85 (m, 1H), 7.24–7.37 (m, 5H), 7.63*
(d, 1/3H, J=9.6 Hz); 13C NMR (125 MHz, CDCl3, both
rotamers) d ppm: 11.7, 14.1, 14.2, 14.4, 14.8, 14.9, 21.71,
21.75, 22.7, 23.2, 23.4, 24.3, 24.5, 24.7, 26.0, 26.3, 26.4,
29.3, 29.55, 29.59, 29.66, 29.72, 31.1, 31.9, 34.3, 35.6,
35.7, 36.8, 37.5, 37.7, 40.8, 41.3, 52.6, 54.8, 56.1, 56.6,
59.2, 66.18, 66.21, 69.5, 73.4, 73.5, 74.4, 75.6, 118.8,
118.9, 127.7, 127.91, 127.93, 128.2, 128.5, 128.6, 131.3,
131.4, 136.8, 137.3, 169.3, 169.6, 170.71, 170.75, 170.8,
171.5, 176.8, 178.1; IR (CHCl3) cmÀ1: 3431, 2961, 2929,
2856, 1746, 1672, 1622, 1511, 1503, 1466; HRMS calcd
for C40H68N3O7 (M+H)+ calcd 702.5057, found
702.5071; [a]2D4=À63.0 (c 1.25, CHCl3).
0.82 (d, 2H, J=7.0 Hz), 0.84–1.00 (m, 10H), 0.96 (d,
2H, J=6.6 Hz), 1.04–1.20 (m, 1H), 1.08* (d, 1H, J=6.6
Hz), 1.23 (d, 2H, J=7.0 Hz), 1.22–1.46 (m, 10H), 1.59–
1.70 (m, 1H), 1.77–1.84 (m, 3H), 1.89–2.00 (m, 1H),
2.69–2.76 (m, 2/3H), 2.80* (s, 1H), 2.86–2.92* (m, 1/
3H), 2.95 (s, 2H), 3.30 (s, 3H), 3.59–3.64 (m, 5/3H), 3.80
(br, 1H), 3.80–3.87 (m, 1H), 4.47 (dd, 2/3H, J=4.4, 8.7
Hz), 4.58* (dd, 1/3H, J=4.3, 9.5 Hz), 4.67* (dd, 1/3H,
J=4.3, 10.4 Hz), 4.74–4.79 (m, 1H), 5.09 (dd, 1H,
J=5.6, 10.0 Hz), 5.13 and 5.25 (AB type d’s, each 2/3H,
J=12.3 Hz), 5.16* and 5.21* (AB type d’s, each 1/3H,
J=12.3 Hz), 6.63 (d, 2/3H), 6.82–6.85 (m, 1H), 7.31–
7.40 (m, 5H), 7.63* (d, 1/3H, J=9.6 Hz); 13C NMR
(125 MHz, CDCl3, both rotamers): d ppm: 11.7, 14.06,
14.12, 14.3, 14.78, 14.85, 21.7, 21.8, 22.6, 23.2, 23.4,
24.3, 24.5, 24.7, 25.9, 26.3, 26.4, 29.3, 29.7, 31.2, 31.7,
31.8, 34.3, 35.6, 35.8, 36.8, 37.5, 37.6, 40.9, 41.4, 52.6,
56.1, 56.6, 59.1, 59.18, 59.21, 67.3, 67.4, 72.0, 74.4, 75.7,
128.05, 128.12, 128.4, 128.51, 128.55, 128.61, 135.1,
135.2, 169.6, 169.8, 170.7, 170.79, 170.83, 171.5, 176.9,
178.1; IR (CHCl3) cmÀ1: 3431, 2962, 2931, 1746, 1672,
1624, 1502, 1460, 1118; HRMS calcd for C34H58N3O7
(M+H)+ 620.4275, found 620.4270; [a]2D4 À70.5 (c 1.05,
CHCl3).
4.1.34. (2R,3R)-3-Hydroxy-2-methylnonanoyl-L-Leu-L-
allo-Ile-L-Ser(Bn)-OAllyl (20j). 92%, a colorless solid
(mp, 152–153 ꢀC), H NMR (400 MHz, CDCl3) d ppm:
1
4.1.32. (2R,3R)-3-Hydroxy-2-methylnonanoyl-N-Me-L-
Leu-L-Ile-L-Ser(Bn)-OAllyl (20d). 94%,
a
colorless
0.86–0.95 (m, 15H), 1.10–1.30 (m, 8H), 1.22 (d, 3H,
J=7.0 Hz), 1.36–1.51 (m, 4H), 1.54–1.73 (m, 3H), 1.90–
1.99 (m, 1H), 2.29–2.36 (m, 1H), 3.23 (br, 1H), 3.55–
3.64 (m, 1H), 3.67 (dd, 1H, J=3.2, 9.6 Hz), 3.94 (dd,
1H, J=3.6, 9.6 Hz), 4.43–4.48 (m, 1H), 4.46 and 4.55
(AB type d’s, each 1H, J=12.3 Hz), 4.50–4.56 (m, 1H),
4.58–4.68 (m, 2H), 4.78 (dt, 1H, J=3.2, 8.4 Hz), 5.23
(dt, 1H, J=1.2, 10.2 Hz), 5.31 (dt, 1H, J=1.3, 18.8 Hz),
5.81–5.91 (m, 1H), 6.23 (d, 1H, J=7.8 Hz), 6.79 (d, 1H,
J=8.1 Hz), 6.99 (d, 1H, J=8.9 Hz), 7.23–7.38 (m, 5H);
13C NMR (125 MHz, CDCl3) d ppm: 11.8, 14.1, 14.3,
15.4, 21.8, 22.6, 23.0, 24.9, 25.8, 26.1, 29.3, 31.8, 35.5,
37.8, 40.5, 46.6, 52.2, 52.5, 56.5, 66.2, 69.5, 73.3, 74.2,
118.8, 127.8, 128.0, 128.5, 131.5, 137.2, 169.5, 171.1,
172.2, 176.0; IR (KBr) cmÀ1: 3284, 2958, 2930, 2973,
1745, 1640, 1547, 1397, 1212, 1114; HRMS calcd for
C35H58N3O7 (M+H)+ calcd 632.4275, found; 632.4274.
amorphous foam, 1H NMR [400 MHz, CDCl3, two
rotamers (major/minor=2/1)] d ppm: 0.86–0.97 (m,
15H), 1.07* (d, 1H, J=6.6 Hz), 1.08–1.17 (m, 1H), 1.23
(d, 2H, J=6.9 Hz), 1.27–1.36 (m, 7H), 1.37–1.54 (m,
4H), 1.57–1.64 (m, 1H), 1.66–2.00 (m, 3H), 2.69–2.76
(m, 2/3H), 2.80* (s, 1H), 2.83–2.87* (m, 1/3H), 2.95 (s,
2H), 3.30* (br s, 1/3H), 3.62* (br, 1H), 3.67 (dd, 2/3H,
J=3.4, 9.5 Hz), 3.68* (dd, 1/3H, J=3.0, 9.5 Hz), 3.70–
3.84 (br, 1H), 3.93 (dd, 1H, J=3.0, 9.5 Hz), 4.31 (dd, 2/
3H, J=5.9, 8.4 Hz), 4.42* (dd, 1/3H, J=5.9, 9.2 Hz),
4.44–4.56* (m, 2/3H), 4.48 and 4.56 (AB type d’s, each
2/3H, J=12.1 Hz), 4.59–4.68 (m, 2H), 4.72–4.78 (m,
1H), 5.10 (dd, 2/3H, J=5.5, 10.2 Hz), 5.22–5.26 (m,
1H), 5.28–5.33 (m, 1H), 5.81–5.91 (m, 1H), 6.60 (d, 2/
3H, J=8.2 Hz), 6.75–6.81 (m, 1H), 7.24–7.38 (m, 5H),
7.73* (d, 1/3H, J=9.3 Hz); 13C NMR (125 MHz,
CDCl3, both rotamers) d ppm: 11.27, 11.32, 14.1, 14.7,
14.8, 15.4, 15.6, 21.67, 21.73, 22.6, 23.3, 23.4, 24.3,
24.48, 24.53, 24.7, 25.9, 29.3, 29.5, 31.1, 31.7, 31.8, 34.3,
35.6, 35.76, 35.80, 37.4, 37.5, 40.8, 41.4, 52.6, 57.4, 57.9,
59.1, 66.2, 69.5, 73.4, 73.5, 74.4, 75.6, 118.8, 118.9,
127.7, 127.8, 127.9, 128.1, 128.5, 128.6, 131.3, 131.5,
136.9, 137.3, 169.3, 169.5, 170.48, 170.51, 170.6, 171.2,
176.8, 178.0; IR (KBr) cmÀ1: 3274, 2957, 2931, 2875,
1742, 1642, 1618, 1540, 1455, 1115; HRMS calcd for
C36H60O7N3 (M+H)+ calcd 646.4431, found 646.4436.
Anal. calcd for C36H59N3O7: C, 66.95; H, 9.21; N, 6.51;
Found: C, 66.75; H, 9.24; N, 6.40; [a]2D3 À72.6 (c 1.10,
CHCl3).
.
Anal. calcd for C35H57N3O7 1/4H2O: C, 66.06; H, 9.11;
N, 6.60, found: C, 66.07; H, 9.05; N, 6.68; [a]2D4 À25.9 (c
1.07, CHCl3).
4.1.35. (2R,3R)-3-Hydroxy-2-methylnonanoyl-N-Et-L-Leu-
L-allo-Ile-L-Ser(Bn)-OAllyl (20k). 28%, a slightly yel-
low oil, 1H NMR [400 MHz, CDCl3, two rotamers
(major/minor=2.8/1)] d ppm: 0.80 (d, 3/4H, J=6.7 Hz),
0.86–0.99 (m, 45/4H), 0.95 (d, 9/4H, J=6.6 Hz), 0.98*
(d, 3/4H, J=6.6 Hz), 1.06* (d, 3/4H, J=6.6 Hz), 1.12–
1.44 (m, 69/4H), 1.47–1.65 (m, 2H), 1.79–1.90 (m, 1H),
1.93–1.99 (m, 1H), 2.66–2.74 (m, 3/4H), 2.83–2.86* (m,
1/4H), 3.27–3.36 (m, 1H), 3.48–3.57 (m, 1H), 3.65–3.81
(m, 3H), 3.92 (dd, 1H, J=3.2, 9.5 Hz), 4.43–4.69 (m,
4H), 4.47 and 4.56 (AB type d’s, each 1H, J=12.3 Hz),
4.74–4.79 (m, 1H), 5.23 (dd, 1H, J=1.4, 9.8 Hz), 5.29
(dd, 1H, J=1.4, 18.8 Hz), 5.81–5.92 (m, 1H), 6.67 (d, 3/
4H, J=8.2 Hz), 6.83* (d, 1/4H, J=7.9 Hz), 7.23–7.36
4.1.33. (2R,3R)-3-Hydroxy-2-methylnonanoyl-N-Me-L-
Leu-L-allo-Ile-L-Ser(Me)-OBn (20i). 100%, a slightly
1
yellow oil, H NMR [400 MHz, CDCl3, two rotamers
(major/mino=2.0/1)] d ppm: 0.77* (d, 1H, J=6.8 Hz),

350
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
(m, 5H), 7.50–7.55 (m, 1H); 13C NMR (125 MHz,
CDCl3, both rotamers) d ppm: 11.75, 11.80, 14.09,
14.12, 14.2, 14.3, 14.5, 15.1, 15.47, 15.52, 21.8, 22.3,
22.57, 22.64, 22.9, 23.3, 24.1, 24.3, 25.0, 25.5, 25.6, 26.4,
29.1, 29.36, 29.44, 31.6, 31.69, 31.74, 31.8, 32.7, 34.4,
35.3, 37.5, 37.6, 37.8, 39.7, 41.3, 42.0, 52.6, 56.3, 56.6,
59.7, 66.17, 66.21, 69.58, 69.63, 73.4, 73.6, 74.5, 75.4,
77.5, 118.7, 118.9, 127.7, 127.9, 128.0, 128.2, 128.5,
128.6, 131.4, 131.5, 136.8, 137.3, 169.3, 169.6, 171.1,
51.8, 52.6, 56.3, 56.7, 57.4, 66.21, 66.23, 69.5, 69.6, 73.4,
73.5, 74.1, 75.2, 118.8, 118.9, 127.7, 127.9, 128.1, 128.5,
128.6, 131.4, 131.5, 136.9, 137.3, 169.4, 169.6, 170.1,
170.76, 170.78, 171.6, 175.7, 177.0; IR (CHCl3) cmÀ1
:
3431, 2959, 2932, 2873, 1746, 1671, 1641, 1502, 1457,
1255; HRMS calcd for C34H53N3O7Na (M+Na)+
calcd 652.3938, found 652.3925; [a]2D4 À57.2 (c 1.04,
CHCl3).
171.2, 171.4, 172.1, 176.2, 178.6; IR (CHCl3) cmÀ1
:
4.1.38. (3R)-Hydroxynonanoyl-N-Me-L-Leu-L-allo-Ile-L-
1
3432, 2961, 2932, 2873, 1746, 1671, 1614, 1503, 1385,
1252; HRMS calcd for C37H61N3O7Na (M+Na)+
682.4407, found 682.4400; [a]2D4 À32.4 (c 1.28, CHCl3).
Ser(Bn)-OAllyl (20n). 100%, a colorless oil, H NMR
[500 MHz, CDCl3, two rotamers (major/minor=2.6/1)]
d ppm: 0.80* (d, 6/7H, J=6.8 Hz), 0.85 (d, 15/7H,
J=6.8 Hz), 0.84–0.97 (m, 9H), 0.95 (d, 3H, J=6.8 Hz),
1.07–1.50 (m, 13H), 1.55–1.76 (m, 2H), 1.94–2.00 (m,
1H), 2.30* (d, 2/7H, J=13.7 Hz), 2.37 (dd, 5/7H,
J=8.8, 16.6 Hz), 2.52 (dd, 5/7H, J=2.9, 16.6 Hz), 2.68*
(dd, 2/7H, J=9.8, 13.7 Hz), 2.81* (s, 6/7H), 2.91 (s, 15/
7H), 3.68 (dd, 5/7H, J=2.9, 9.8 Hz), 3.76* (dd, 2/7H,
J=2.9, 9.8 Hz), 3.93 (dd, 5/7H, J=2.9, 9.8 Hz), 3.95*
(dd, 2/7H, J=2.9, 9.8 Hz), 3.98–4.07 (m, 1H), 4.18–4.31
(m, 1H), 4.43–4.44 (m, 1H), 4.44 and 4.51* (AB type
d’s, each 2/7H, J=11.7 Hz), 4.48 and 4.56 (AB type d’s,
each 5/7H, J=11.7 Hz), 4.59–4.69 (m, 2H), 4.73–4.77
(m, 1H), 5.14 (dd, 1H, J=6.8, 9.8 Hz), 5.23–5.33 (m,
2H), 5.82–5.91 (m, 1H), 6.64 (d, 5/7H, J=7.8 Hz), 6.75
(d, 5/7H, J=8.8 Hz), 6.97* (d, 2/7H, J=7.8 Hz), 7.23–
7.37 (m, 5H), 7.42* (d, 2/7H, J=9.8 Hz); 13C NMR
(125 MHz, CDCl3, both rotamers) d ppm: 11.67, 11.74,
14.05, 14.09, 14.31, 14.34, 21.9, 22.1, 22.59, 22.63, 23.0,
23.3, 24.4, 24.9, 25.2, 25.6, 26.3, 26.5, 29.2, 29.6, 31.1,
31.6, 31.7, 31.8, 36.1, 36.4, 36.9, 37.5, 37.6, 37.9, 40.0,
52.6, 54.8, 56.3, 59.9, 66.19, 66.22, 68.1, 69.5, 69.7, 71.4,
73.4, 73.7, 118.78, 118.84, 127.7, 127.9, 128.2, 128.4,
128.5, 128.7, 131.3, 131.4, 137.3, 169.3, 169.5, 170.4,
4.1.36. (2R,3R)-3-Hydroxy-2-methylnonanoyl-L-Pro-L-
allo-Ile-L-Ser(Bn)-OAllyl (20l). 96%, a colorless solid
(mp, 81–82 ꢀC), H NMR [400 MHz, CDCl3, two rota-
1
mers (major/minor=2/1)] d ppm: 0.86–0.94 (m, 9H),
1.10–1.20 (m, 1H), 1.23 (d, 3H, J=7.1 Hz), 1.28–1.41
(m, 8H), 1.41–1.55 (m, 3H), 1.70–1.95 (m, 2H), 1.96–
2.06 (m, 2H), 2.09–2.16 (m, 1H), 2.29–2.34* (m, 1/3H),
2.37–2.45 (m, 1H), 2.61 (qu, 2/3H, J=7.0 Hz), 3.51–
3.61 (m, 2H), 3.63–3.71 (m, 2H), 3.77* (br t, 1/3H,
J=9.1 Hz), 3.92 (dd, 2/3H, J=3.4, 9.5 Hz), 3.93–3.97*
(m, 1/3H), 4.35–4.45 (m, 1H), 4.45 (dd, 2/3H, J=4.2,
8.3 Hz), 4.48 and 4.55 (AB type d’s, each 2/3H, J=12.2
Hz), 4.55–4.59 (m, 2/3H), 4.60–4.67 (m, 2H), 4.68–4.71
(m, 1H), 4.73–4.79 (m, 1H), 5.21–5.26 (m, 1H), 5.31 (dq,
1H, J=1.3, 16.9 Hz), 5.81–5.92 (m, 1H), 6.75 (d, 2/3H,
J=8.3 Hz), 6.78* (d, 1/3H, J=8.7 Hz), 7.14* (d, 1/3H,
J=9.1 Hz), 7.27–7.37 (m, 5H), 7.50 (d, 2/3H, J=8.4
Hz); 13C NMR (125 MHz, CDCl3, both rotamers) d
ppm: 11.4, 11.7, 14.06, 14.2, 14.6, 15.1, 22.59, 22.62,
25.1, 25.2, 25.8, 26.3, 27.0, 29.3, 31.7, 31.8, 33.8, 35.3,
37.0, 37.1, 43.0, 46.1, 46.4, 47.5, 52.60, 52.67, 52.68,
56.7, 57.2, 59.5, 61.4, 66.1, 66.2, 69.4, 69.6, 73.3, 73.4,
73.9, 74.4, 76.5, 118.7, 118.9, 127.68, 127.78, 127.82,
128.0, 128.5, 131.3, 131.5, 137.1, 137.4, 169.3, 169.7,
171.0, 171.8, 172.1, 176.2, 177.1; IR (KBr) cmÀ1: 3385,
3296, 2959, 2927, 1734, 1643, 1626, 1552, 1454, 1203;
HRMS calcd for C34H53N3O7Na (M+Na)+ calcd
638.3781, found 638.3765. Anal. calcd for C34H53N3O7:
C, 66.31; H, 8.68; N, 6.82, found: C, 65.95; H, 8.42; N,
6.74; [a]2D5 À64.9 (c 1.07, CHCl3).
170.7, 170.8, 171.1, 172.8, 174.0; IR (CHCl3) cmÀ1
:
3431, 2961, 2931, 2873, 1746, 1673, 1627, 1502, 1273,
1103; HRMS calcd for C35H57N3O7Na (M+Na)+
654.4094, found 654.4089; [a]2D5 À57.9 (c 1.26, CHCl3).
4.1.39. (2R,3R)-3-Boc-amino-2-methylnonanoyl-N-Mel-
L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (20o). 85%, a color-
1
less oil, H NMR (400 MHz, CDCl3) d ppm: 0.83 (d,
3H, J=7.0 Hz), 0.83–0.92 (m, 9H), 0.95 (d, 3H, J=6.7
Hz), 1.06–1.14 (m, 1H), 1.18 (d, 3H, J=7.0 Hz), 1.17–
1.49 (m, 11H), 1.44 (s, 9H), 1.54–1.61 (m, 1H), 1.67–
1.81 (m, 2H), 1.92–1.98 (m, 1H), 2.93 (s, 3H), 2.93–2.99
(m, 1H), 3.63–3.69 (m, 1H), 3.67 (dd, 1H, J=3.1, 9.5
Hz), 3.93 (dd, 1H, J=3.1, 9.4 Hz), 4.46 (dt, 1H, J=2.9,
8.1 Hz), 4.48 and 4.57 (AB type d’s, each 1H, J=12.3
Hz), 4.59–4.69 (m, 2H), 4.75 (m, 1H), 5.18–5.21 (m,
1H), 5.24 (d, 1H, J=10.0 Hz), 5.30 (d, 1H, J=17.4 Hz),
5.81–5.91 (m, 1H), 5.94 (d, 1H, J=9.6 Hz), 6.58 (d, 1H,
J=8.1 Hz), 6.69 (d, 1H, J=8.7 Hz), 7.28–7.37 (m, 5H);
13C NMR (125 MHz, CDCl3) d ppm: 11.7, 14.0, 14.2,
14.5, 21.6, 22.58, 22.63, 23.3, 24.6, 26.3, 26.6, 28.3, 28.4,
29.2, 30.5, 31.6, 31.8, 34.1, 35.5, 37.6, 38.4, 52.6, 53.56,
53.63, 56.1, 66.2, 69.5, 73.3, 78.7, 118.8, 127.6, 127.7,
128.0, 128.5, 131.4, 137.3, 156.3, 169.6, 170.68, 170.73,
177.2; IR (CHCl3) cmÀ1: 3430, 2962, 2931, 2873, 1745,
1697, 1675, 1626, 1500, 1165; HRMS calcd for
C41H68N4O8Na (M+Na)+ 767.4935, found 767.4934;
[a]2D4 À73.4 (c 1.13, CHCl3).
4.1.37. (2R,3R)-3-Hydroxy-2-methylnonanoyl-L-homo-
Pro-L-allo-Ile-L-Ser(Bn)-OAllyl (20m). 91%, a colorless
1
oil, H NMR [400 MHz, CDCl3, two rotamers (major/
minor=3/2)] d ppm: 0.82–0.96 (m, 9H), 1.07* (d, 6/5H,
J=6.9 Hz), 1.10–1.55 (m, 15H), 1.23 (d, 9/5H, J=7.2
Hz), 1.65–1.86 (m, 2H), 1.91–2.02 (m, 1H), 2.26 (br d, 3/
5H, J=13.4 Hz), 2.49–2.58 (m, 1H), 2.69–2.80 (m, 1H),
3.07–3.14* (m, 2/5H), 3.66–3.69 (m, 8/5H), 3.76* (dt, 2/
5H, J=2.2, 8.3 Hz), 3.83 (br d, 3/5H, J=13.4 Hz),
3.90–3.95 (m, 7/5H), 4.44–4.58 (m, 3H), 4.58–4.67 (m,
3H), 4.73–4.80 (m, 1H), 5.22–5.27 (m, 2H), 5.31 (dd,
1H, J=1.3, 15.6 Hz), 5.81–5.92 (m, 1H), 6.60 (d, 3/5H,
J=8.2 Hz), 6.75 (d, 3/5H, J=8.5 Hz), 6.89* (d, 2/5H,
J=8.1 Hz), 7.24–7.38 (m, 27/5H); 13C NMR (125 MHz,
CDCl3, both rotamers) d ppm: 11.6, 11.8, 14.09, 14.12,
14.4, 14.6, 14.8, 15.0, 20.3, 21.1, 22.62, 22.66, 24.8, 25.0,
25.3, 25.5, 26.0, 26.4, 26.56, 26.64, 29.3, 29.4, 31.6,
31.77, 31.85, 34.3, 35.3, 37.4, 37.7, 39.9, 40.7, 41.7, 43.7,

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
351
4.1.40. (2R,3S)-3-Hydroxy-2-methylnonanoyl-N-Me-L-
Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (20p). 80%, a colorless
oil, H NMR [500 MHz, CDCl3, two rotamers (major/
66.2, 68.7, 69.5, 73.3, 76.4, 80.2, 118.7, 127.7, 127.9,
128.4, 131.5, 137.3, 155.7, 169.0, 169.6, 170.3, 170.7,
170.8, 174.4; IR (CHCl3) cmÀ1: 3430, 2959, 2930, 2858,
1744, 1677, 1636, 1499, 1471, 1256; HRMS calcd for
C57H99N5O12SiNa (M+Na)+ calcd 1096.6941, found
1096.6949. Anal. calcd for C57H99N5O12Si H2O: C,
62.66; H, 9.32; N, 6.41, found: C, 62.89; H, 9.28; N,
6.59; [a]2D4 À51.9 (c 1.02, CHCl3).
1
minor=5.7/1)] d ppm: 0.80* (d, 3/7H, J=6.8 Hz), 0.83
(d, 18/7H, J=6.8 Hz), 0.86–0.93 (m, 10H), 0.95 (d, 18/
7H, J=6.8 Hz), 0.96* (d, 3/7H, J=6.8 Hz), 1.09 (d, 3/
7H, J=6.8 Hz), 1.15 (d, 18/7H, J=6.8 Hz), 1.23–1.35
(m, 9H), 1.43–1.59 (m, 3H), 1.68–1.73 (m, 2H), 1.93–
2.01 (m, 1H), 2.64 (dq, 6/7H, J=2.9, 6.8 Hz), 2.78* (s,
3/7H), 2.94 (s, 18/7H), 3.15–3.20* (m, 1/7H), 3.43–3.48*
(m, 1/7H), 3.67 (dd, 6/7H, J=3.4, 9.3 Hz), 3.74* (dd, 1/
7H, J=2.9, 9.8 Hz), 3.81–3.85 (m, 1H), 3.93 (dd, 1H,
J=3.4, 9.3 Hz), 4.10–4.13 (m, 6/7H), 4.44–4.49 (m, 1H),
4.47 and 4.56 (AB type d’s, each 1H, J=12.7 Hz), 4.62–
4.67 (m, 2H), 4.74–4.77 (m, 1H), 5.11 (dd, 1H, J=6.8,
8.8 Hz), 5.23 (dd, 1H, J=2.0, 8.8 Hz), 5.30 (dd, 1H,
J=2.0, 17.6 Hz), 5.82–5.91 (m, 1H), 6.63 (d, 6/7H,
J=7.8 Hz), 6.73 (d, 6/7H, J=8.8 Hz), 6.77* (d, 1/7H,
J=8.8 Hz), 7.25–7.37 (m, 5H), 7.64* (d, 1/7H, J=9.8
Hz), 13C NMR (125 MHz, CDCl3, both rotamers) d
ppm: 9.9, 11.7, 14.1, 14.2, 22.0, 22.6, 23.0, 24.9, 26.0,
26.3, 29.3, 30.8, 31.8, 34.0, 35.8, 37.4, 39.9, 52.6, 54.4,
56.1, 66.2, 69.5, 71.7, 73.4, 118.8, 127.7, 127.9, 128.5,
128.6, 131.5, 137.3, 169.6, 170.5, 170.7, 178.8; IR
(CHCl3) cmÀ1: 3432, 2961, 2933, 2873, 1746, 1673, 1616,
1502, 1466, 1106; HRMS calcd for C36H60N3O7
(M+H)+ 646.4431, found 646.4429; [a]2D5 À78.3 (c 1.00,
CHCl3).
.
4.1.42. (2R,3R)-3-[Boc-L-allo-Thr(TBS)-GlyO]-2-methyl-
nonanoyl-N-Me-L-Leu-L-Ile-L-Ser(Bn)-OAllyl (21d). 95%,
a colorless amorphous foam, 1H NMR (400 MHz,
CDCl3) d ppm: 0.05 (s, 3H), 0.08 (s, 3H), 0.81–0.92 (m,
21H), 0.94 (d, 3H, J=6.7 Hz), 1.11 (d, 3H, J=6.8 Hz),
1.18 (d, 3H, J=6.4 Hz), 1.05–1.35 (m, 7H), 1.37–1.48
(m, 4H), 1.44 (s, 9H), 1.55–1.65 (m, 3H), 1.71–1.79 (m,
1H), 1.88–1.94 (m, 1H), 2.99 (s, 3H), 3.08 (qu, 1H,
J=6.8 Hz), 3.66 (dd, 1H, J=3.1, 9.5 Hz), 3.77 (dd, 1H,
J=4.2, 8.4 Hz), 3.92 (dd, 1H, J=3.0, 9.5 Hz), 4.08–4.19
(m, 3H), 4.27 (dd, 1H, J=6.0, 8.3 Hz), 4.47 and 4.56
(AB type d’s, each 1H, J=12.1 Hz), 4.60–4.68 (m, 2H),
4.74 (dt, 1H, J=3.1, 8.1 Hz), 5.08 (d, 1H, J=7.4 Hz),
5.11–5.16 (m, 2H), 5.23 (dd, 1H, J=1.2, 10.4 Hz), 5.30
(dt, 1H, J=1.2, 17.0 Hz), 5.81–5.90 (m, 1H), 6.63 (d,
1H, J=8.3 Hz), 6.68–6.71 (m, 2H), 7.26–7.36 (m, 5H);
13C NMR (125 MHz, CDCl3) d ppm: À5.0, À4.6, 11.2,
12.3, 14.0, 15.5, 17.9, 19.5, 21.8, 22.5, 23.2, 23.5, 24.5,
24.8, 25.0, 25.7, 28.3, 29.1, 29.9, 30.5, 31.6, 35.8, 37.0,
39.9, 41.1, 52.6, 54.3, 57.5, 66.2, 68.8, 69.5, 73.3, 76.3,
80.2, 118.7, 127.7, 127.8, 128.4, 131.5, 137.3, 155.7,
4.1.41. (2R,3R)-3-[Boc-L-allo-Thr(TBS)-GlyO]-2-methyl-
tridecanoyl-N-Me-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (21c).
To a solution of 20c (202 mg, 0.287 mmol) and 15a
(136 mg, 0.348 mmol) in CH2Cl2 (1.5 mL), were added
DMAP (38 mg, 0.31 mmol), CSA (33 mg, 0.14 mmol)
followed by DIPC (68 mL, 0.43 mmol) at room tem-
perature. The reaction mixture was stirred at the same
temperature. Moreover, 15a and other reagents were
sometimes added in order to finish the reaction. The
mixture was concentrated and diluted with hexane and
EtOAc. The mixture was filtered and the filtrate was
washed with 10% aqueous citric acid, saturated
NaHCO3, aqueous solution and brine. The organic
layer was dried over Na2SO4, filtered and evaporated.
The residue was purified by column chromatography
with a gradient elution system, a 3:1–2:1 mixture of
hexane and EtOAc used as eluents to give 21c (299 mg,
169.0, 169.5, 170.3, 170.5, 170.6, 174.4; IR (KBr) cmÀ1
:
3315, 2959, 2931, 2859, 1749, 1655, 1526, 1464, 1388,
1196; HRMS calcd for C53H92N5O12Si (M+H)+ calcd
1018.6512, found 1018.6509. Anal. calcd for
.
C53H91N5O12Si H2O: C, 61.42; H, 9.04; N, 6.76; Found:
C, 61.65; H, 9.04; N, 6.95; [a]2D4 À55.3 (c 1.11, CHCl3).
4.1.43.
(2R,3R)-3-[Boc-L-Thr(TBS)-GlyO]-2-methyl-
nonanoyl-N-Me-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (21e).
94%, a colorless amorphous foam, 1H NMR (400 MHz,
CDCl3) d ppm: 0.06 (s, 3H), 0.09 (s, 3H), 0.81–0.90 (m,
21H), 0.94 (d, 3H, J=6.6 Hz), 1.06–1.14 (m, 7H), 1.18–
1.35 (m, 9H), 1.39–1.50 (m, 1H), 1.47 (s, 9H), 1.54–1.65
(m, 3H), 1.70–1.77 (m, 1H), 1.95–2.01 (m, 1H), 2.99 (s,
3H), 3.08 (qu, 1H, J=7.0 Hz), 3.66 (dd, 1H, J=3.1, 9.5
Hz), 3.88–3.95 (br m, 1H), 3.92 (dd, 1H, J=3.0, 9.5 Hz),
4.05 (dd, 1H, J=5.0, 18.7 Hz), 4.13 (br d, 1H, J=6.1
Hz), 4.44 (dd, 2H, J=4.5, 8.7 Hz), 4.47 and 4.56 (AB
type d’s, each 1H, J=12.1 Hz), 4.58–4.68 (m, 2H), 4.74
(dt, 1H, J=3.0, 8.1 Hz), 5.13–5.18 (br m, 2H), 5.23 (dt,
1H, J=1.3, 10.3 Hz), 5.30 (dq, 1H, J=1.3, 17.6 Hz),
5.37 (br d, 1H, J=7.2 Hz), 5.81–5.89 (m, 1H), 6.65 (d,
1H, J=8.3 Hz), 6.69 (br s, 1H), 7.11 (br t, 1H, J=4.5
Hz), 7.26–7.36 (m, 5H); 13C NMR (100 MHz, CDCl3) d
ppm: À5.1, À4.7, 11.7, 12.3, 14.0, 14.3, 17.9, 19.3, 21.8,
22.6, 23.2, 24.8, 25.1, 25.7, 26.3, 28.3, 29.1, 30.0, 30.4,
31.7, 35.8, 37.2, 40.0, 41.3, 52.7, 54.2, 56.2, 59.6, 66.2,
68.2, 69.5, 73.3, 76.3, 77.5, 80.1, 118.8, 127.7, 127.9,
128.5, 131.5, 137.4, 155.9, 169.0, 169.6, 170.4, 170.8,
174.5; IR (KBr) cmÀ1: 3317, 2958, 2932, 2860, 1748,
1719, 1654, 1522, 1199, 1170; HRMS calcd for
C53H92N5O12Si (M+H)+ calcd 1018.6512, found
1
0.278 mmol, 97%) as a colorless amorphous foam. H
NMR (400 MHz, CDCl3) d ppm: 0.05 (s, 3H), 0.08 (s,
3H), 0.81–0.93 (m, 21H), 0.95 (d, 3H, J=6.7 Hz), 1.10
(d, 3H, J=6.8 Hz), 1.18 (d, 3H, J=6.0 Hz), 1.19–1.39
(m, 18H), 1.38–1.50 (m, 1H), 1.44 (s, 9H), 1.46–1.65 (m,
3H), 1.71–1.80 (m, 1H), 1.95–2.01 (m, 1H), 2.99 (s, 3H),
3.08 (qu, 1H, J=6.9 Hz), 3.67 (dd, 1H, J=3.0, 9.5 Hz),
3.78 (dd, 1H, J=4.1, 8.5 Hz), 3.92 (dd, 1H, J=3.0, 9.5
Hz), 4.08–4.42 (m, 3H), 4.42–4.45 (m, 1H), 4.47 and
4.56 (AB type d’s, each 1H, J=12.0 Hz), 4.58–4.68 (m,
2H), 4.74 (dt, 1H, J=3.1, 8.1 Hz), 5.08–5.16 (m, 3H),
5.23 (dt, 1H, J=1.3, 10.2 Hz), 5.31 (dt, 1H, J=1.3, 17.6
Hz), 5.81–5.91 (m, 1H), 6.66–6.72 (m, 3H), 7.26–7.36
(m, 5H); 13C NMR (125 MHz, CDCl3) d ppm: À5.0,
À4.7, 11.6, 12.2, 14.1, 14.2, 17.9, 19.5, 21.8, 22.6, 23.2,
24.8, 25.1, 25.5, 25.7, 26.3, 28.3, 29.3, 29.5, 29.6, 29.9,
30.5, 31.9, 35.8, 37.2, 39.9, 41.1, 52.6, 54.3, 56.2, 60.8,
.
1018.6515. Anal. calcd for C53H91N5O12Si H2O: C,

352
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
61.42; H, 9.04; N, 6.76; Found: C, 61.23; H, 8.94; N,
7.13; [a]2D4 À55.4 (c 1.08, CHCl3).
3H, J=6.6 Hz), 1.11 (d, 3H, J=6.8 Hz), 1.15 (d, 3H,
J=6.5 Hz), 1.20 (d, 3H, J=6.3 Hz), 1.22–1.33 (m, 9H),
1.45 (s, 9H), 1.58–1.65 (m, 2H), 1.71–1.79 (m, 1H),
1.94–2.01 (m, 1H), 2.99 (s, 3H), 3.08 (qu, 1H, J=6.9
Hz), 3.35 (s, 3H), 3.61–3.68 (m, 2H), 3.81–3.93 (m, 2H),
4.11–4.13 (m, 1H), 4.27 (br t, 1H, J=6.7 Hz), 4.42–4.44
(m, 1H), 4.47 and 4.56 (AB type d’s, each 1H, J=12.0
Hz), 4.59–4.68 (m, 2H), 4.72–4.76 (m, 1H), 5.11–5.20
(m, 3H), 5.23 (dd, 1H, J=1.3, 11.4 Hz), 5.30 (dd, 1H,
J=1.3, 17.6 Hz), 5.81–5.91 (m, 1H), 6.66 (d, 1H, J=8.1
Hz), 6.71 (br d, 1H, J=6.3 Hz), 6.84 (br t, 1H, J=4.8
Hz), 7.26–7.36 (m, 5H); 13C NMR (125 MHz, CDCl3) d
ppm: 11.6, 12.5, 14.0, 14.2, 15.3, 21.8, 22.5, 23.2, 23.4,
24.8, 25.0, 26.3, 28.3, 29.1, 30.1, 30.6, 31.6, 35.8, 37.2,
39.9, 41.3, 42.3, 52.6, 54.4, 56.2, 56.7, 66.2, 69.5, 73.3,
76.3, 77.5, 118.7, 127.7, 127.9, 128.4, 131.5, 137.3, 169.6,
170.5, 170.8, 174.5; IR (CHCl3) cmÀ1: 3433, 2963, 2933,
2873, 1745, 1674, 1500, 1369, 1252, 1164; HRMS calcd
for C48H80N5O12 (M+H)+ 918.5803, found 918.5811;
[a]2D5 À64.1 (c 1.21, CHCl3).
4.1.44. (2R,3R)-3-[Boc-L-Thr(TBS)-GlyO]-2-methylnon-
anoyl-N-Me-L-Leu-L-Ile-L-Ser(Bn)-OAllyl (21f). 96%, a
colorless amorphous foam, 1H NMR (400 MHz,
CDCl3) d ppm: 0.06 (s, 3H), 0.09 (s, 3H), 0.83–0.90 (m,
21H), 0.94 (d, 3H, J=6.3 Hz), 1.10–1.14 (m, 7H), 1.18–
1.37 (m, 9H), 1.38–1.54 (m, 2H), 1.46 (s, 9H), 1.55–1.66
(m, 2H), 1.70–1.78 (m, 1H), 1.88–1.94 (m, 1H), 2.99 (s,
3H), 3.07 (qu, 1H, J=6.9 Hz), 3.66 (dd, 1H, J=3.1, 9.5
Hz), 3.87–3.94 (br m, 1H), 3.92 (dd, 1H, J=3.0, 9.5 Hz),
4.01–4.13 (m, 2H), 4.28 (dd, 1H, J=6.2, 8.5 Hz), 4.41–
4.44 (m, 1H), 4.47 and 4.55 (AB type d’s, each 1H,
J=12.2 Hz), 4.59–4.66 (m, 2H), 4.74 (dt, 1H, J=3.1, 8.2
Hz), 5.13–5.16 (m, 2H), 5.23 (dd, 1H, J=1.3, 10.2 Hz),
5.30 (dq, 1H, J=1.3, 15.2 Hz), 5.37 (br d, 1H, J=7.4
Hz), 5.81–5.90 (m, 1H), 6.65 (d, 1H, J=8.2 Hz), 6.68 (br
d, 1H, J=8.4 Hz), 7.10 (br t, 1H, J=4.5 Hz), 7.25–7.36
(m, 5H); 13C NMR (125 MHz, CDCl3) d ppm: À5.1,
À4.7, 11.2, 12.3, 14.0, 15.5, 17.8, 19.3, 21.8, 22.6, 23.2,
24.5, 24.8, 25.0, 25.7, 28.3, 29.1, 30.4, 31.7, 35.8, 37.0,
39.9, 41.3, 52.6, 54.1, 57.5, 59.6, 66.2, 68.2, 69.5, 73.3,
76.3, 77.5, 80.1, 118.8, 127.7, 127.9, 128.4, 131.5, 137.3,
155.8, 169.0, 169.5, 170.4, 170.5, 170.6, 174.5; IR
(CHCl3) cmÀ1: 3426, 2959, 2931, 2860, 1745, 1713, 1674,
1495, 1368, 1257; HRMS calcd for C53H92N5O12Si
(M+H)+ calcd 1018.6512, found 1018.6502. Anal.
4.1.47. (2R,3R)-3-[Boc-L-Thr(TBS)-GlyO]-2-methylnon-
anoyl-N-Me-L-Leu-L-allo-Ile-L-Ser(Me)-OBn (21i). 91%,
a slightly yellow oil, 1H NMR (400 MHz, CDCl3) d
ppm: 0.05 (s, 3H), 0.08 (s, 3H), 0.80 (d, 3H, J=7.1 Hz),
0.83–0.92 (m, 9H), 0.87 (s, 9H), 0.95 (d, 3H, J=6.6 Hz),
1.10 (d, 3H, J=6.8 Hz), 1.18 (d, 3H, J=6.1 Hz), 1.20–
1.31 (m, 10H), 1.40–1.48 (m, 2H), 1.44 (s, 9H), 1.59–
1.66 (m, 2H), 1.70–1.77 (m, 1H), 1.92–1.99 (m, 1H), 2.99
(s, 3H), 3.08 (t, 1H, J=6.9 Hz), 3.29 (s, 3H), 3.59 (dd,
1H, J=3.2, 9.4 Hz), 3.78 (dd, 1H, J=4.5, 19.0 Hz), 3.83
(dd, 1H, J=3.2, 9.4 Hz), 4.08–4.19 (m, 3H), 4.44 (dd,
1H, J=4.5, 8.6 Hz), 4.73–4.77 (m, 1H), 5.08–5.15 (m,
3H), 5.14 and 5.25 (AB type d’s, each 1H, J=12.3 Hz),
6.63 (d, 1H, J=8.2 Hz), 6.70–6.71 (m, 2H), 7.30–7.39
(m, 5H); 13C NMR (125 MHz, CDCl3) d ppm: À5.0,
À4.6, 11.6. 12.3, 14.0, 14.1, 14.2, 17.9, 19.5, 21.9, 22.5,
22.6, 23.2, 24.8, 25.0, 25.7, 26.3, 28.3, 29.1, 29.9, 30.5,
31.6, 35.8, 37.2, 39.9, 41.1, 52.6, 54.3, 56.1, 59.2, 67.3,
68.7, 72.0, 76.3, 76.5, 80.2, 128.1, 128.4, 128.5, 135.3,
155.7, 169.0, 169.8, 170.3, 170.7, 170.8, 174.4; IR
(CHCl3) cmÀ1: 3431, 2959, 2931, 2859, 1744, 1676, 1500,
1257, 1163, 1118; HRMS calcd for C51H90N5O12Si
(M+H)+ 992.6355, found 992.6365; [a]2D5 À56.8 (c 1.07,
CHCl3).
.
calcd for C53H91N5O12Si 1/2H2O: C, 61.96; H, 9.03; N,
6.82; Found: C, 61.97; H, 8.91; N, 7.14; [a]2D4 À55.6 (c
1.14, CHCl3).
4.1.45. (2R,3R)-3-(Boc-L-Abu-GlyO)-2-methylnonanoyl-
N-Me-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (21g). 90%, a
colorless amorphous foam, 1H NMR (400 MHz,
CDCl3, major conformer) d ppm: 0.83–0.96 (m, 19H),
1.11 (d, 3H, J=6.8 Hz), 1.15 (t, 3H, J=6.6 Hz), 1.20–
1.40 (m, 5H), 1.42–1.47 (m, 3H), 1.45 (s, 9H), 1.58–1.68
(m, 2H), 1.72–1.84 (m, 2H), 1.86–1.92 (m, 1H), 1.95–
1.99 (m, 1H), 2.99 (s, 3H), 3.03–3.10 (m, 1H), 3.67 (dd,
1H, J=3.0, 9.5 Hz), 3.84–3.87 (m, 1H), 3.92 (dd, 1H,
J=3.2, 9.5 Hz), 4.06–4.11 (m, 2H), 4.42–4.44 (m, 1H),
4.46 and 4.57 (AB type d’s, each 1H, J=12.0 Hz), 4.62–
4.64 (m, 2H), 4.72–4.76 (m, 1H), 5.02 (d, 1H, J=8.1
Hz), 5.10 (dd, 1H, J=6.0, 9.5 Hz), 5.18 (dt, 1H, J=2.9,
7.8 Hz), 5.23 (dd, 1H, J=1.4, 10.3 Hz), 5.30 (dd, 1H,
J=1.4, 16.9 Hz), 5.81–5.91 (m, 1H), 6.61 (br s, 1H),
6.66 (d, 1H, J=8.2 Hz), 6.70 (br d, 1H, J=7.2 Hz),
7.26–7.36 (m, 5H); 13C NMR (125 MHz, CDCl3, both
rotamers) d ppm: 9.9, 11.7, 12.5, 14.0, 14.3, 21.8, 22.6,
23.2, 23.5, 24.8, 25.0, 25.8, 26.3, 28.3, 29.1, 30.2, 30.7,
31.6, 35.9, 37.2, 40.0, 41.3, 42.3, 52.7, 54.6, 55.7, 56.2,
66.2, 69.5, 73.3, 76.4, 76.6, 80.1, 118.8, 127.7, 127.9,
128.5, 131.5, 137.4, 155.6, 169.2, 169.6, 170.6, 172.1,
174.6; IR (CHCl3) cmÀ1: 3433, 2964, 2933, 2874, 1744,
1677, 1500, 1369, 1255, 1163; HRMS calcd for
C47H78N5O11 (M+H)+ 888.5698, found 888.5708; [a]2D4
À68.7 (c 1.51, CHCl3).
4.1.48. (2R,3R)-3-[Boc-L-allo-Thr(TBS)-GlyO]-2-methyl-
nonanoyl-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (21j). 100%,
ꢀ
1
a colorless solid (mp128–130 C), H NMR (400 MHz,
CDCl3) d ppm: 0.05 (s, 3H), 0.07 (s, 3H), 0.85–0.94 (m,
24H), 1.13 (d, 3H, J=7.2 Hz), 1.18 (d, 3H, J=6.1 Hz),
1.15–1.29 (m, 10H), 1.39–1.47 (m, 1H), 1.43 (s, 9H),
1.55–1.67 (m, 4H), 1.90–1.96 (m, 1H), 2.60 (qu, 1H,
J=6.9 Hz), 3.67 (dd, 1H, J=3.1, 9.5 Hz), 3.76 (dd, 1H,
J=4.0, 17.9 Hz), 3.93 (dd, 1H, J=3.1, 9.5 Hz), 4.14–
4.20 (m, 2H), 4.24 (dd, 1H, J=6.5, 17.9 Hz), 4.46–4.53
(m, 2H), 4.47 and 4.56 (AB type d’s, each 1H, J=12.3
Hz), 4.59–4.68 (m, 2H), 4.75 (dt, 1H, J=3.1, 8.2 Hz),
5.03 (q, 1H, J=5.7 Hz), 5.21 (br, 1H), 5.23 (dd, 1H,
J=1.3, 10.3 Hz), 5.28–5.33 (m, 1H), 5.81–5.91 (m, 1H),
6.34 (br s, 1H), 6.67 (br, 2H), 7.13 (br, 1H), 7.25–7.36
(m, 5H); 13C NMR (125 MHz, CDCl3) d ppm: À5.0,
À4.6, 11.8, 14.0, 14.2, 17.9, 19.6, 19.8, 21.9, 22.6, 23.0,
4.1.46. (2R,3R)-3-[Boc-L-allo-Thr(Me)-GlyO]-2-methyl-
nonanoyl-N-Me-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (21h).
86%,
a
colorless amorphous foam, 1H NMR
(400 MHz, CDCl3) d ppm: 0.81–0.91 (m, 12H), 0.95 (d,

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
353
24.8, 25.1, 25.7, 26.1, 28.3, 29.1, 31.3, 31.6, 38.0, 40.6,
41.4, 44.7, 51.9, 52.7, 56.5, 66.2, 68.7, 69.5, 73.3, 77.5,
77.6, 80.2, 118.8, 127.7, 127.9, 128.5, 131.5, 137.3, 155.7,
169.1, 169.5, 170.6, 170.9, 172.3, 173.0; IR (CHCl3)
cmÀ1: 3294, 2959, 2932, 2860, 1744, 1642, 1540, 1367,
1201, 1115; HRMS calcd for C52H90N5O12Si (M+H)+
calcd 1004.6356, found 1004.6356. Anal. calcd for
C52H89N5O12Si: C, 62.18; H, 8.93; N, 6.97, found: C,
61.84; H, 8.69; N, 7.05; [a]2D4 À7.2 (c 1.06, CHCl3).
C51H85N5O12SiNa (M+Na)+ calcd 1010.5861, found
.
1010.5862. Anal. calcd for C51H85N5O12Si 1/2H2O: C,
61.42; H, 8.69; N, 7.02, found: C, 61.31; H, 8.74; N,
7.16; [a]2D5 À38.4 (c 1.07, CHCl3).
4.1.51. (2R,3R)-3-[Boc-L-allo-Thr(TBS)-GlyO]-2-methyl-
nonanoyl-L-homo-Pro-L-allo-Ile-L-Ser(Bn)-OAllyl (21m).
97%, a colorless amorphous foam, 1H NMR (400 MHz,
CDCl3) d ppm: 0.05 (s, 3H), 0.08 (s, 3H), 0.84 (d, 3H,
J=7.1 Hz), 0.85–0.93 (m, 15H), 1.12 (d, 3H, J=7.0 Hz),
1.18 (d, 3H, J=6.0 Hz), 1.19–1.39 (m, 10H), 1.45 (s,
9H), 1.40–1.52 (m, 2H), 1.57–1.72 (m, 2H), 1.74–1.85
(m, 3H), 1.92–1.98 (m, 1H), 2.23 (br d, 1H, J=13.2 Hz),
3.09–3.16 (m, 2H), 3.66 (dd, 1H, J=3.1, 9.5 Hz), 3.83
(dd, 1H, J=4.2, 18.4 Hz), 3.92 (dd, 1H, J=3.0, 9.5 Hz),
4.02 (br d, 1H, J=13.2 Hz), 4.08–4.20 (m, 3H), 4.42
(dd, 1H, J=4.5, 8.4 Hz), 4.47 and 4.54 (AB type d’s,
each 1H, J=12.1 Hz), 4.60–4.68 (m, 2H), 4.74 (dt, 1H,
J=3.1, 8.2 Hz), 5.07–5.15 (m, 2H), 5.20 (br d, 1H, J=4.8
Hz), 5.24 (dd, 1H, J=1.3, 10.5 Hz), 5.30 (dq, 1H,
J=1.4, 16.7 Hz), 5.81–5.91 (m, 1H), 6.63 (d, 1H, J=8.2
Hz), 6.71 (d, 1H, J=5.4 Hz), 6.74 (d, 1H, J=8.3 Hz),
7.26–7.36 (m, 5H); 13C NMR (125 MHz, CDCl3) d ppm:
À5.0, À4.6, 11.8, 12.1, 14.0, 14.1, 14.4, 17.9, 19.6, 20.3,
22.6, 22.7, 25.2, 25.6, 25.7, 25.9, 26.4, 28.3, 29.0, 29.8,
31.6, 37.4, 39.4, 41.2, 43.6, 52.2, 52.6, 56.4, 66.2, 68.8,
69.5, 73.3, 76.4, 118.8, 127.7, 127.9, 128.5, 131.5, 137.4,
155.7, 169.1, 169.6, 170.4, 170.8, 171.0, 173.6; IR (KBr)
cmÀ1: 3317, 2957, 2931, 2858, 1747, 1657, 1525, 1366,
1196, 835; HRMS calcd for C52H87N5O12SiNa
(M+Na)+ calcd 1024.6018, found 1024.6019. Anal.
4.1.49. (2R,3R)-3-(Boc-L-allo-Thr(TBS)-GlyO)-2-methyl-
nonanoyl-N-Et-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (21k).
1
94%, colorless amorphous foam, H NMR (400 MHz,
CDCl3) d ppm: 0.05 (s, 3H), 0.08 (s, 3H), 0.84–0.94 (m,
12H), 0.87 (s, 9H), 0.93 (d, 3H, J=6.6 Hz), 1.13 (d, 3H,
J=6.9 Hz), 1.17 (d, 3H, J=6.2 Hz), 1.17–1.32 (m, 12H),
1.38–1.44 (m, 3H), 1.44 (s, 9H), 1.59–1.79 (m, 3H),
1.96–2.03 (m, 1H), 3.00–3.07 (m, 1H), 3.26–3.35 (m,
1H), 3.53–3.60 (m, 1H), 3.67 (dd, 1H, J=3.4, 9.5 Hz),
3.77 (dd, 1H, J=4.0, 18.4 Hz), 3.90 (dd, 1H, J=3.0, 9.5
Hz), 4.09–4.20 (m, 3H), 4.40 (dd, 1H, J=4.4, 8.2 Hz),
4.46 and 4.56 (AB type d’s, each 1H, J=12.2 Hz), 4.61–
4.64 (m, 2H), 4.72–4.75 (m, 1H), 4.95–5.00 (m, 1H),
5.06–5.09 (m, 1H), 5.11–5.16 (m, 1H), 5.23 (dd, 1H,
J=1.3, 10.3 Hz), 5.30 (dd, 1H, J=1.3, 17.7 Hz), 5.81–
5.91 (m, 1H), 6.69 (d, 1H, J=8.1 Hz), 6.73 (t, 1H,
J=4.9 Hz), 6.96 (d, 1H, J=8.3 Hz), 7.26–7.36 (m, 5H);
13C NMR (125 MHz, CDCl3) d ppm: À5.0, À4.6, 11.7,
13.3, 13.7, 14.01, 14.04, 14.4, 16.0, 17.9, 19.5, 19.6, 22.3,
22.5, 22.6, 22.9, 24.8, 24.9, 25.2, 25.69, 25.72, 26.4, 28.3,
28.8, 29.1, 30.2, 31.56, 31.64, 31.9, 36.7, 36.9, 41.0, 41.1,
52.7, 56.5, 65.5, 66.2, 68.8, 69.5, 73.3, 76.9, 118.7, 127.7,
127.8, 128.4, 131.5, 137.4, 168.9, 169.6, 170.3, 170.9,
171.7, 175.2; IR (CHCl3) cmÀ1: 3431, 2960, 2931, 2860,
1745, 1676, 1500, 1385, 1257, 1163; HRMS calcd for
C54H93N5O12SiNa (M+Na)+ 1054.6488, found
1054.6432; [a]2D4 À50.2 (c 1.06, CHCl3).
.
calcd for C52H87N5O12Si 1/2H2O: C, 61.75; H, 8.77; N,
6.92, found: C, 61.57; H, 8.43; N, 6.83; [a]2D4 À50.5 (c
1.06, CHCl3).
4.1.52. (3R)-3-[Boc-L-allo-Thr(TBS)-GlyO]-nonanoyl-N-
Me-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (21n). 90%, a col-
1
orless amorphous foam, H NMR (400 MHz, CDCl3,
4.1.50. (2R,3R)-3-[Boc-L-allo-Thr(TBS)-GlyO]-2-methyl-
nonanoyl-L-Pro-L-allo-Ile-L-Ser(Bn)-OAllyl (21l). 95%,
a colorless amorphous foam, 1H NMR (400 MHz,
CDCl3) d ppm: 0.05 (s, 3H), 0.08 (s, 3H), 0.85–0.90 (m,
18H), 1.12 (d, 3H, J=7.1 Hz), 1.18 (d, 3H, J=6.3 Hz),
1.21–1.42 (m, 9H), 1.44 (s, 9H), 1.50–1.61 (m, 1H),
1.71–1.77 (m, 1H), 1.80–1.90 (m, 2H), 1.98–2.04 (m,
2H), 2.08–2.21 (m, 1H), 2.38–2.42 (m, 1H), 2.93 (qu,
1H, J=7.2 Hz), 3.57 (dt, 1H, J=3.0, 8.5 Hz), 3.61–3.68
(m, 1H), 3.68 (dd, 1H, J=3.4, 9.5 Hz), 3.76 (dd, 1H,
J=4.2, 18.4 Hz), 3.90 (dd, 1H, J=3.1, 9.5 Hz), 4.08–
4.18 (m, 3H), 4.40 (dd, 1H, J=4.6, 8.3 Hz), 4.47 and
4.55 (AB type d’s, each 1H, J=12.2 Hz), 4.58–4.68 (m,
3H), 4.74 (dt, 1H, J=3.2, 8.1 Hz), 5.07 (br d, 1H, J=6.6
Hz), 5.16 (dt, 1H, J=2.8, 8.2 Hz), 5.22 (dt, 1H, J=1.3,
10.4 Hz), 5.30 (dq, 1H, J=1.4, 17.4 Hz), 5.81–5.91 (m,
1H), 6.70 (t, 1H, J=4.2 Hz), 6.74 (d, 1H, J=8.1 Hz),
7.26–7.36 (m, 5H), 7.45 (d, 1H, J=8.3 Hz); 13C NMR
(125 MHz, CDCl3) d ppm: À5.0, À4.6, 11.7, 13.0, 14.1,
14.3, 17.9, 19.6, 22.6, 24.8, 25.1, 25.7, 26.3, 27.1, 28.3,
29.1, 30.4, 31.7, 37.0, 41.2, 41.5, 47.5, 52.7, 56.9, 59.6,
66.1, 68.8, 69.6, 73.3, 76.6, 77.5, 118.7, 127.7, 127.8,
128.4, 131.5, 137.5, 155.7, 168.8, 169.7, 170.4, 171.0,
171.3, 173.7; IR (CHCl3) cmÀ1: 3431, 2959, 2931, 2860,
1745, 1714, 1676, 1500, 1256, 1162; HRMS calcd for
major conformer) d ppm: 0.03 (s, 3H), 0.06 (s, 3H),
0.81–1.00 (m, 21H), 0.92 (d, 3H, J=6.6 Hz), 1.14 (d,
3H, J=6.2 Hz), 1.18–1.42 (m, 10H), 1.42 (s, 9H), 1.58–
1.77 (m, 5H), 1.98–2.05 (m, 1H), 2.55 (dd, 1H, J=5.1,
15.1 Hz), 2.65 (dd, 1H, J=8.0, 15.1 Hz), 2.90 (s, 3H),
3.66 (dd, 1H, J=3.0, 9.5 Hz), 3.77 (dd, 1H, J=4.0, 18.4
Hz), 3.93 (dd, 1H, J=2.9, 9.5 Hz), 4.11–4.21 (m, 2H),
4.25 (dd, 1H, J=6.6, 18.4 Hz), 4.40 (dd, 1H, J=5.9, 8.8
Hz), 4.46 and 4.57 (AB type d’s, each 1H, J=12.2 Hz),
4.59–4.68 (m, 2H), 4.78–4.81 (m, 1H), 5.16 (dd, 1H,
J=6.0, 9.5 Hz), 5.22 (dd, 1H, J=1.3, 10.3 Hz), 5.20–
5.36 (m, 3H), 5.80–5.90 (m, 1H), 6.68 (d, 1H, J=8.3
Hz), 6.77 (d, 1H, J=8.8 Hz), 6.97 (dd, 1H, J=4.1, 5.8
Hz), 7.25–7.36 (m, 5H); 13C NMR (125 MHz, CDCl3,
both rotamers) d ppm: À5.0, À4.7, 11.5, 14.0, 14.5, 17.9,
19.6, 21.9, 22.6, 23.2, 24.9, 25.2, 25.6, 25.7, 26.2, 28.2,
28.3, 29.0, 31.0, 31.6, 34.3, 36.1, 36.9, 38.7, 41.2, 52.7,
54.6, 56.8, 66.2, 68.9, 69.6, 72.2, 73.4, 77.5, 118.7, 127.7,
127.9, 128.5, 131.5, 137.4, 155.6, 169.6, 169.7, 170.5,
170.75, 170.81, 170.9; IR (CHCl3) cmÀ1: 3431, 3365,
2959, 2931, 1743, 1676, 1499, 1368, 1256, 1163; HRMS
calcd for C52H89N5O12SiNa (M+Na)+ 1026.6175,
.
found 1026.6206; anal. calcd for C52H89N5O12Si 3/
2H2O: C, 60.55; H, 8.99; N, 6.79, found: C 60.31; H,
8.68; N, 6.97; [a]2D6 À33.1 (c 1.06, CHCl3).

354
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
4.1.53. (2R,3R)-3-[Boc-N-Me-L-allo-Thr(TBS)-GlyNH]-
2-methylnonanoyl-N-Me-L-Leu-L-allo-Ile-L-Ser(Bn)-
OAllyl (21o). 99%, a colorless amorphous foam, 1H
NMR [400 MHz, CD3OD, two rotamers (major/
minor=4.5/1)] d ppm: 0.05 (s, 3H), 0.08 (s, 3H), 0.85–
0.98 (m, 21H), 0.96 (d, 3H, J=6.5 Hz), 1.05–1.45 (m,
14H), 1.08 (d, 3H, J=6.8 Hz), 1.17 (d, 3H, J=5.9 Hz),
1.45 (s, 9H), 1.78–1.88 (m, 1H), 1.90–1.93 (m, 1H),
2.95* (s, 1/2H), 3.04 (s, 5/2H), 3.04–3.16 (m, 1H), 3.68–
3.74 (m, 1H), 3.74 (dd, 1H, J=3.8, 9.8 Hz), 3.89 (dd,
1H, J=4.5, 9.8 Hz), 3.95–4.00 (m, 2H), 4.03–4.05 (m,
1H), 4.09–4.20 (m, 1H), 4.45–4.48 (m, 1H), 4.49 and
4.57 (AB type d’s, each 5/6H, J=12.1 Hz), 4.50* and
4.58* (AB type d’s, each 1/6H, J=11.9 Hz), 4.62–4.64
(m, 2H), 4.70 (t, 5/6H, J=4.5 Hz), 4.73* (t, 1/6H,
J=4.3 Hz), 5.20 (dd, 1H, J=2.7, 11.6 Hz), 5.24–5.26
(m, 1H), 5.31 (dd, 1H, J=2.7, 18.9 Hz), 5.85–5.96 (m,
1H), 7.25–7.35 (m, 5H); 13C NMR (125 MHz, CD3OD,
both rotamers) d ppm: À4.8, À4.4, 12.0, 12.1, 13.5, 14.4,
14.9, 15.1, 18.8, 20.3, 22.1, 23.3, 23.65, 23.71, 23.8, 25.9,
26.0, 26.3, 27.4, 27.5, 28.8, 30.1, 30.3, 31.5, 32.2, 32.9,
37.4, 38.9, 40.5, 43.8, 53.2, 54.3, 57.6, 58.1, 62.4, 67.0,
69.1, 69.7, 70.4, 70.5, 74.2, 80.8, 118.7, 128.8, 128.9, 129.4,
132.4, 133.2, 139.1, 170.9, 171.1, 172.9, 173.5, 178.1; IR
(CHCl3) cmÀ1: 3429, 3347, 2960, 2931, 2859, 1668, 1501,
1368, 1257, 1162; HRMS calcd for C53H92N6O11SiNa
(M+Na)+ 1039.6491, found 1039.6516. Anal. calcd for
C53H92N6O11Si: C, 62.57; H, 9.11; N, 8.26, found: C
62.46; H, 8.84; N, 8.09; [a]2D4 À50.7 (c 1.29, CHCl3).
1.71–1.78 (m, 2H), 1.93–2.01 (m, 1H), 3.00 (s, 3H), 3.11
(qu, 1H, J=6.4 Hz), 3.19 (s, 3H), 3.53 (d, 1H, J=17.2
Hz), 3.66 (dd, 1H, J=3.0, 9.5 Hz), 3.85 (dd, 1H, J=6.2,
8.1 Hz), 3.91 (dd, 1H, J=3.3, 9.5 Hz), 4.43 (dd, 1H,
J=4.5, 8.7 Hz), 4.46 and 4.55 (AB type d’s, each 1H,
J=12.1 Hz), 4.53–4.58 (m, 1H), 4.58–4.67 (m, 3H), 4.73
(dt, 1H, J=3.1, 8.2 Hz), 5.03 (br q, 1H, J=5.7 Hz), 5.15
(dd, 1H, J=6.0, 9.7 Hz), 5.21–5.24 (m, 2H), 5.30 (dq,
1H, J=1.3, 16.7 Hz), 5.80–5.90 (m, 1H), 6.64 (d, 1H,
J=8.2 Hz), 6.70 (d, 1H, J=8.7 Hz), 7.25–7.36 (m,
5H); 13C NMR (125 MHz, CDCl3, both rotamers) d
ppm: À5.1, À5.0, À4.9, À4.8, 11.5, 11.7, 14.0, 14.1, 14.2,
17.8, 20.6, 20.9, 21.88, 21.94, 22.55, 22.64, 23.1,
23.2, 24.7, 25.4, 25.7, 26.3, 26.9, 28.27, 28.32, 29.1, 29.4,
30.4, 31.56, 31.63, 35.1, 35.7, 35.8, 37.1, 37.2, 39.9, 49.7,
52.6, 54.2, 55.3, 56.2, 66.2, 69.5, 71.1, 73.3, 75.7, 77.5,
79.6, 118.7, 127.7, 127.9, 128.4, 131.5, 137.4, 155.1,
168.5, 169.6, 170.76, 170.79, 172.6, 174.4; IR (CHCl3)
cmÀ1: 3433, 2960, 2931, 2860, 1746, 1710, 1650, 1501,
1258, 1164; HRMS calcd for C54H94N5O12Si (M+H)+
calcd 1032.6669, found 1032.6680. Anal. calcd for
.
C54H93N5O12Si 1/5H2O: C, 62.60; H, 9.09; N, 6.76,
found: C, 62.33; H, 8.64; N, 6.82; [a]2D7 À52.8 (c 1.17,
CHCl3).
4.1.56. General procedure of deprotections and macro-
lactamization: compound 22c. To a solution of 21c (287
mg, 0.267 mmol) in THF (2.0 mL) was added a mixture
of AcOH (110 mL, 1.92 mmol) and TBAF (1.0 M THF
solution, 1.35 mL, 1.35 mmol) at 0 ꢀC. The solution was
stirred at the same temperature for 2 h and at room
temperature for an additional 16 h. This reaction mix-
ture was diluted with EtOAc and washed with 5% aqu-
eous KHSO4 solution, saturated NaHCO3 aqueous
solution and brine. The resulting organic layer was
dried over Na2SO4, filtered and evaporated. The residue
was purified by column chromatography with a gradient
elution system, a 2:1–1:1 mixture of hexane and EtOAc
used as eluents to give a alcohol (233 mg, 0.242 mmol,
91%) as a colorless amorphous foam. This compound
(231 mg, 0.241 mmol) and morpholine (63 mL, 0.72
mmol) were dissolved in THF (2.5 mL). To this solution
was added Pd(PPh3)4 (14.2 mg) and the reaction mix-
ture was stirred at room temperature for 18 h. The
mixture was concentrated and purified by column chro-
matography with EtOAc and then a 10:1 mixture of
EtOAc and CH3OH used as an eluents to give a acid
(208 mg, 0.226 mmol, 94%) as a slightly yellow amor-
phous foam. To a solution of the acid (150.4 mg, 163
mmol) in CH2Cl2 (2.0 mL) was added TFA (0.4 mL) at
room temperature. After being stirred at the same tem-
perature, the solvents were removed in vacuo. The resi-
due was dissolved in CH2Cl2 and reconcentrated
repeatedly to remove excess TFA. The crude products
was dried under reduced pressure to afford a TFA salt
(169 mg) and which was used for macrocyclization
without further purification. This salt (51.0 mg, 49.3
mmol) was dissolved in THF (27 mL) and the solution
was slowly added to a suspension of HATU (208 mg,
0.547 mmol) and DIEA (185 mL) in THF (27.6 mL) for
over 10 h at 0 ꢀC with a syringe pump under high dilu-
tion condition. After being stirred at the same tempera-
ture for an additional 7 h and at room temperature for 1
4.1.54. (2R,3S)-3-(Boc-L-Thr(TBS)-GlyO)-2-methylnon-
anoyl-N-Me-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl (21p). 82%,
a yellow amorphous foam, 1H NMR (500 MHz, CDCl3)
d ppm: 0.05 (s, 3H), 0.08 (s, 3H), 0.83 (d, 3H, J=6.8
Hz), 0.85–0.90 (m, 9H), 0.87 (s, 9H), 0.96 (d, 3H, J=6.8
Hz), 1.12 (d, 3H, J=6.8 Hz), 1.17–1.33 (m, 10H), 1.20
(d, 3H, J=5.9 Hz), 1.42–1.64 (m, 4H), 1.44 (s, 9H),
1.72–1.78 (m, 1H), 1.95–2.00 (m, 1H), 2.91–2.95 (m,
1H), 2.94 (s, 3H), 3.67 (dd, 1H, J=2.9, 9.8 Hz), 3.90–
3.95 (m, 2H), 4.07–4.16 (m, 3H), 4.44 (dd, 1H, J=4.9,
8.8 Hz) 4.47 and 4.56 (AB type d’s, each 1H, J=12.2
Hz), 4.60–4.67 (m, 2H), 4.75 (dt, 1H, J=2.9, 7.8 Hz),
5.09 (d, 1H, J=7.8 Hz), 5.16 (dd, 1H, J=5.9, 9.8 Hz),
5.23 (dd, 1H, J=2.0, 10.8 Hz), 5.25–5.27 (m, 1H), 5.30
(dd, 1H, J=2.0, 17.6 Hz), 5.82–5.90 (m, 1H), 6.66 (d,
1H, J=7.8 Hz), 6.71–6.77 (m, 2H), 7.27–7.36 (m, 5H),
13C NMR (125 MHz, CDCl3) d ppm: À5.1, À4.6, 11.7,
14.0, 14.1, 14.2, 17.9, 19.7, 21.8, 22.5, 23.2, 24.7, 25.2,
25.7, 26.3, 28.3, 29.0, 30.5, 31.6, 32.4, 35.7, 37.2, 40.4,
41.3, 52.6, 54.0, 56.2, 66.2, 68.8, 69.5, 73.3, 77.5, 118.8,
127.7, 127.9, 128.4, 131.5, 137.3, 169.0, 169.6, 170.5,
170.7, 170.8, 175.0; IR (KBr) cmÀ1: 3312, 2958, 2931,
2859, 1747, 1653, 1525, 1389, 1198, 1111; HRMS calcd
for C53H92N5O12Si (M+H)+ 1018.6512, found
1018.6519; [a]2D4 À52.6 (c 1.41, CHCl3).
4.1.55. (2R,3R)-3-[Boc-N-Me-L-allo-Thr(TBS)-GlyO]-2-
methylnonanoyl-N-Me-L-Leu-L-allo-Ile-L-Ser(Bn)-OAllyl
(21q). 100%, a colorless amorphous foam, 1H NMR
(400 MHz, CDCl3, major conformer) d ppm: À0.04 (s,
3H), 0.04 (s, 3H), 0.80–0.90 (m, 21H), 0.92–0.95 (m,
3H), 1.08 (d, 3H, J=6.8 Hz), 1.20–1.30 (m, 11H), 1.22
(d, 3H, J=6.1 Hz), 1.42 (s, 9H), 1.56–1.70 (m, 2H),

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
355
day, the mixture was evaporated, diluted with 3:1 mix-
ture of CH3OH and EtOAc and filtered. The filtrate was
concentrated, dissolved in EtOAc and washed with 1%
aqueous HCl solution, saturated NaHCO3 aqueous
solution and then brine. The organic layer was dried
over Na2SO4, filtered and evaporated. The residue was
purified by preparative HPLC (column: Develosil ODS-
HG5 (10 mmÂ250 mm); wavelength: 210 nm; flow rate:
4.0 mL/min) with an 89:11 mixture of CH3OH and 1%
aqueous triethylammmonium acetate as an eluent to
give 22c [13.0 mg, 16.2ꢀmmol, 33% (two steps)] as a col-
orless solid (mp72–74 C). 1H NMR [500 MHz, CDCl3,
two rotamers (major/minor=3.5/1)] d ppm: 0.85–0.98
(m, 15H), 1.10 (d, 12/5H, J=6.9 Hz), 1.16* (d, 3/5H,
J=7.0 Hz), 1.18–1.67 (m, 24H), 1.72–1.76 (m, 1H),
1.82–1.92 (m, 1H), 1.99–2.04 (m, 1H), 2.08–2.16 (m,
1H), 2.79* (s, 3/5H), 3.08–3.14 (m, 1H), 3.17 (s, 12/5H),
3.62 (dd, 4/5H, J=3.9, 17.2 Hz), 3.68 (br s, 4/5H), 3.80*
(dd, 1/5H, J=3.7, 18.5 Hz), 3.84 (dd, 1H, J=4.7, 10.0
Hz), 3.89 (dd, 4/5H, J=5.5, 10.0 Hz), 3.93* (dd, 1/5H,
J=5.9, 10.0 Hz), 4.01* (q, 1/5H, J=6.0 Hz), 4.11–4.22
(m, 9/5H), 4.29 (dd, 4/5H, J=4.9, 9.7 Hz), 4.34* (dd, 1/
5H, J=4.6, 9.1 Hz), 4.37–4.42 (m, 4/5H), 4.45* (t, 1/5H,
J=7.0 Hz), 4.54 and 4.55 (AB type d’s, each 1H,
J=11.8 Hz), 4.77* (dd, 1/5H, J=3.8, 9.5 Hz), 4.84* (d,
1/5H, J=10.8 Hz), 5.10–5.14 (m, 4/5H), 6.75* (d, 1/5H,
J=4.3 Hz), 6.81–6.88 (m, 9/5H), 7.17* (d, 1/5H, J=8.6
Hz), 7.24* (dd, 1/5H, J=3.9, 7.3 Hz), 7.31–7.38 (m,
5H), 7.62 (br d, 4/5H, J=4.7 Hz), 7.87 (br s, 4/5H); 13C
NMR (125 MHz, CDCl3, both rotamers) d ppm: 11.69,
11.74, 11.78, 14.1, 14.6, 14.8, 15.1, 19.8, 19.9, 21.8, 22.5,
22.6, 23.04, 23.11, 24.2, 24.7, 25.2, 26.3, 26.9, 27.1, 28.8,
29.27, 29.35, 29.41, 29.51, 29.54, 29.57, 31.2, 31.9, 36.8,
37.5, 38.1, 38.4, 39.4, 40.4, 40.7, 55.4, 55.7, 56.2, 56.8,
57.3, 58.0, 59.2, 67.65, 67.69, 68.0, 73.4, 73.8, 78.1,
127.9, 128.2, 128.4, 128.6, 128.7, 136.6, 137.0, 169.0,
169.2, 169.8, 170.0, 170.8, 170.9, 171.5, 172.3, 172.8,
173.8, 174.3, 176.7; IR (KBr) cmÀ1: 3328, 2958, 2926,
2856, 1738, 1656, 1542, 1466, 1377, 1201; HRMS calcd
for C43H71N5O9Na (M+Na)+ calcd 824.5150, found
J=7.7 Hz), 7.04 (s, 3/4H), 7.23* (d, 1/4H, J=8.7 Hz),
7.30–7.37 (m, 21/4H), 7.59 (dd, 3/4H, J=4.1, 8.0 Hz),
7.74 (br d, 3/4H, J=4.0 Hz); 13C NMR (100 MHz,
CDCl3, both rotamers) d ppm: 11.87, 11.91, 14.2, 15.1,
16.3, 16.4, 19.7, 19.9, 21.9, 22.68, 22.70, 23.2, 24.3,
24.85, 24.89, 25.0, 25.3, 26.4, 28.9, 29.1, 29.3, 29.5, 29.8,
31.4, 31.7, 31.8, 36.6, 37.7, 38.1, 38.5, 39.5, 39.6, 40.7,
41.0, 41.1, 55.4, 57.2, 57.9, 58.0, 58.9, 59.2, 67.5, 67.65,
67.70, 68.0, 73.4, 73.9, 77.1, 77.2, 78.1, 127.7, 128.0,
128.3, 128.5, 128.6, 136.9, 168.7, 169.1, 169.5, 169.8,
170.8, 171.2, 171.7, 172.5, 173.4, 173.6, 176.2; IR (KBr)
cmÀ1: 3326, 2959, 2929, 2872, 1738, 1656, 1540, 1465,
1202, 1115; HRMS calcd for C39H64N5O9 (M+H)+
calcd 746.4704, found 746.4704.. Anal. calcd for
.
C39H63N5O9 H2O: C, 61.31; H, 8.58; N, 9.17, found: C,
61.69; H, 8.47; N, 9.05; [a]2D6 À6.6 (c 1.02, CHCl3).
4.1.58. 22-epi-24-OBn-globomycin (22e). 50% (2 steps),
ꢀ
1
a colorless solid (mp, 77–79 C), H NMR (500 MHz,
CDCl3, major confomer (major/minor=5.2/1)) d ppm:
0.87–0.98 (m, 15H), 1.12 (d, 3H, J=6.9 Hz), 1.17 (d,
3H, J=6.4 Hz), 1.26–1.31 (m, 8H), 1.36–1.52 (m, 3H),
1.63–1.75 (m, 2H), 1.91–2.08 (m, 3H), 2.19–2.24 (m,
1H), 3.01–3.07 (m, 1H), 3.07–3.13 (m, 1H), 3.19 (s, 3H),
3.53 (dd, 1H, J=3.6, 17.0 Hz), 3.67 (br s, 1H), 3.80 (dd,
1H, J=4.0, 10.0 Hz), 3.90 (dd, 1H, J=4.9, 10.0 Hz),
4.24 (dd, 1H, J=4.1, 7.5 Hz), 4.37 (d, 1H, J=8.8 Hz),
4.50-4.61 (m, 1H), 4.54 and 4.57 (AB type d’s, each 1H,
J=11.9 Hz), 4.70 (q, 1H, J=6.4 Hz), 5.32–5.36 (m, 1H),
6.35 (d, 1H, J=2.8 Hz), 6.71 (d, 1H, J=8.8 Hz), 7.30–
7.39 (m, 5H), 7.93 (dd, 1H, J=3.6, 9.1 Hz), 8.36 (br s,
1H); 13C NMR (100 MHz, CDCl3, both rotamers) d
ppm: 11.8, 12.0, 12.2, 14.16, 14.25, 14.6, 15.0, 15.2, 19.4,
19.5, 21.8, 22.6, 22.7, 22.8, 23.2, 23.4, 24.0, 25.1, 25.3,
26.5, 27.0, 27.2, 27.3, 29.16, 29.29, 29.36, 29.42, 29.8,
31.5, 31.7, 31.8, 37.6, 38.0, 38.3, 38.5, 39.9, 40.0, 40.7,
41.9, 56.1, 56.2, 56.3, 57.1, 57.8, 58.7, 59.3, 66.0, 66.5,
67.8, 68.1, 69.2, 73.2, 73.4, 73.7, 76.5, 76.6, 78.3, 127.8,
128.0, 128.2, 128.3, 128.5, 136.4, 136.7, 169.1, 169.3,
169.78, 169.82, 170.3, 170.8, 171.3, 172.8, 172.9, 174.3,
174.6, 177.0; IR (KBr) cmÀ1: 3324, 2959, 2930, 2872,
1742, 1655, 1543, 1467, 1203, 1116; HRMS calcd for
C39H64N5O9 (M+H)+ calcd 746.4704, found 764.4690.
.
824.5142. Anal. calcd for C43H71N5O9 1/2H2O: C,
63.68; H, 8.95; N, 8.63, found: C, 63.54; H, 8.79; N,
8.63; [a]2D4=+8.0 (c 1.18, CHCl3).
.
Anal. calcd for C39H63N5O9 H2O: C, 61.31; H, 8.58; N,
4.1.57. 25-epi-24-OBn-globomycin (22d). 40% (two
steps), a colorless solid (mp 72–74 ꢀC), 1H NMR
[500 MHz, CDCl3, two rotamers (major/minor=3.0/1)]
d ppm: 0.88 (t, 9/4H, J=6.7 Hz), 0.90–0.96 (m, 42/4H),
1.00 (d, 9/4H, J=6.8 Hz), 1.10 (d, 9/4H, J=7.0 Hz),
1.16* (d, 3/4H, J=7.0 Hz), 1.18–1.43 (m, 39/4H), 1.22
(d, 9/4H, J=6.0 Hz), 1.47–1.58 (m, 1H), 1.57–1.70 (m,
2H), 1.72–1.76 (m, 2H), 1.87–1.99* (m, 2/4H), 2.02–2.09
(m, 3/4H), 2.10–2.15 (m, 3/4H), 2.80* (s, 3/4H), 3.07–
3.12 (m, 1H), 3.17 (s, 9/4H), 3.60* (d, 1/4H, J=3.5 Hz),
3.69 (dd, 6/4H, J=4.3, 17.2 Hz), 3.81* (dd, 1/4H,
J=3.9, 18.3 Hz), 3.85–3.93* (m, 2/4H), 3.89 (t, 6/4H,
J=5.4 Hz), 3.99* (d, 1/4H, J=4.6 Hz), 4.13–4.23 (m,
12/4H), 4.27 (q, 3/4H, J=5.3 Hz), 4.30–4.35 (m, 5/4H),
4.41* (dd, 1/4H, J=5.4, 8.5 Hz), 4.45* (t, 1/4H, J=7.1
Hz), 4.54 and 4.56 (AB type d’s, each 1H, J=11.7 Hz),
4.62* (dd, 1/4H, J=4.8, 9.3 Hz), 4.85* (d, 1/4H,
J=10.8 Hz), 5.07–5.11 (m, 3/4H), 6.78* (d, 1/4H,
J=4.7 Hz), 6.86* (d, 1/4H, J=9.3 Hz), 6.97 (d, 3/4H,
9.17, found: C, 61.44; H, 8.77; N, 8.94; [a]2D5 +41.3 (c
0.53, CHCl3).
4.1.59. Compound 22f. 41% (two steps), a colorless solid
ꢀ
1
(mp91–93 C), H NMR [500 MHz, CDCl3, two rota-
mers (major/minor=3.8/1)] d ppm: 0.87–0.97 (m, 12H),
1.00* (d, 3/5H, J=6.8 Hz), 1.05 (d, 12/5H, J=6.8 Hz),
1.11–1.14 (m, 27/5H), 1.17* (d, 3/5H, J=6.8 Hz), 1.19–
1.36 (m, 10H), 1.40–1.56 (m, 12/5H), 1.57–1.75 (m, 6/
5H), 1.86–1.99 (m, 6/5H), 2.10–2.16* (m, 1/5H), 2.21–
2.29 (m, 1H), 2.78* (s, 3/5H), 2.83 (br s, 1H), 2.99–3.07
(m, 4/5H), 3.09–3.13* (m, 1/5H), 3.18 (s, 12/5H), 3.55
(dd, 4/5H, J=3.4, 17.1 Hz), 3.70 (br s, 4/5H), 3.75* (dd,
1/5H, J=2.0, 18.6 Hz), 3.83 (dd, 4/5H, J=3.9, 9.8 Hz),
3.90 (dd, 1H, J=4.9, 9.8 Hz), 3.95* (dd, 1/5H, J=3.9,
9.8 Hz), 4.21–4.24 (m, 4/5H), 4.29–4.32* (m, 1/5H),
4.35–4.44 (m, 2H), 4.50 (dd, 4/5H, J=8.8, 17.1 Hz),
4.54–4.56* (m, 1/5H), 4.59 and 4.55 (AB type d’s each
1H, J=11.7 Hz), 4.64–4.69 (m, 6/5H), 4.88* (d, 1/5H,

356
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
J=10.7 Hz), 5.30–5.34 (m, 4/5H), 6.39 (d, 4/5H, J=2.9
Hz), 6.57* (d, 1/5H, J=2.9 Hz), 6.70 (d, 4/5H, J=8.8
Hz), 6.90* (d, 1/5H, J=8.8 Hz), 7.06* (d, 1/5H, J=8.8
Hz), 7.31–7.39 (m, 26/5H), 7.92 (dd, 4/5H, J=3.4, 9.3
Hz), 8.24 (br s, 4/5H); 13C NMR (125 MHz, CDCl3,
both rotamers) d ppm: 11.8, 12.0, 12.2, 14.00, 14.03,
15.1, 16.3, 16.4, 19.2, 19.4, 21.6, 22.50, 22.54, 22.57,
23.1, 23.2, 23.9, 24.6, 24.9, 25.1, 26.3, 29.1, 29.2, 29.3,
31.4, 31.6, 31.7, 37.5, 37.8, 38.1, 38.4, 39.8, 40.68, 40.73,
41.8, 56.1, 56.9, 57.7, 57.9, 58.2, 58.7, 59.2, 65.9, 66.4,
67.8, 68.1, 73.4, 73.7, 76.4, 76.6, 78.3, 127.8, 128.1,
128.2, 128.4, 128.7, 136.6, 136.9, 169.3, 169.5, 169.8,
170.1, 170.7, 171.0, 171.5, 172.6, 173.0, 174.2, 174.3, 177.1;
HRMS calcd for C39H64N5O9 (M+H)+ calcd 746.4704,
3H, J=4.9, 7.8 Hz), 4.35–4.63 (m, 8/3H), 4.52* and
4.59* (AB type d’s, each 1/3H, J=11.7 Hz), 4.55 and
4.60 (AB type d’s, each 2/3H, J=11.7 Hz), 4.80 (dd, 2/
3H, J=3.9, 9.8 Hz), 4.85 (dd, 2/3H, J=3.9, 7.8 Hz),
5.28–5.32 (m, 2/3H), 6.47-6.49 (m, 4/3H), 6.86–6.91 (m,
1H), 6.94–6.96* (m, 1/3H), 7.28–7.39 (m, 14/3H), 7.71
(dd, 2/3H, J=3.9, 8.8 Hz), 8.28 (br s, 1H), 13C NMR
(125 MHz, CDCl3, both rotamers) d ppm: 11.6, 11.8,
14.00, 14.03, 14.75, 14.80, 15.07, 15.10, 21.6, 22.4, 22.57,
22.62, 23.1, 23.2, 24.0, 24.8, 25.1, 26.5, 27.0, 27.1, 28.8,
29.0, 29.2, 29.3, 31.4, 31.6, 31.70, 31.73, 37.4, 37.5, 38.1,
38.5, 39.5, 40.1, 54.8, 55.5, 56.1, 56.2, 56.3, 56.4, 56.67,
56.74, 59.3, 68.0, 73.4, 73.9, 74.9, 75.3, 76.1, 77.5, 78.1,
127.9, 128.18, 128.23, 128.5, 128.60, 128.64, 128.7,
136.6, 136.9, 168.7, 169.0, 169.3, 169.5, 169.66, 169.72,
.
found 746.4707. Anal. calcd for C39H63N5O9 1/3H2O:
C, 62.29; H, 8.53; N, 9.31; Found: C, 62.26; H, 8.60; N,
9.25; [a]2D7,+34.9 (c 1.02, CHCl3).
171.9, 172.6, 173.1, 174.0, 174.6; IR (CHCl3) cmÀ1
:
3349, 2962, 2932, 2872, 1752, 1666, 1545, 1501, 1252,
.
1102. Anal. calcd for C40H65N5O9 2/3H2O: C, 62.23; H,
4.1.60. 22-Deoxy-24-OBn-globomycin (22g). 41% (2
a
colorless amorphous foam, 1H NMR
8.66; N, 9.07, found: C 62.00; H, 8.51; N, 9.03; HRMS
calcd for C40H65N5O9Na (M+Na)+ 782.4680, found
782.4680; [a]2D6 +12.1 (c 1.01, CHCl3).
steps),
[400 MHz, CDCl3, two rotamers (major/minor=2.3/1)]
d ppm: 0.77–1.07 (m, 17H), 1.12* (d, 2H, J=7.0 Hz),
1.16* (d, 1H, J=7.0 Hz), 1.27–1.39 (m, 9H), 1.42–1.58
(m, 3H), 1.60–1.77 (m, 3H), 1.92–2.09 (m, 3H), 2.11–
2.22 (m, 1H), 2.78* (s, 1H), 3.01–3.07 (m, 2/3H), 3.10–
3.16* (m, 1/3H), 3.16 (s, 2H), 3.55 (dd, 2/3H, J=3.4,
17.2 Hz), 3.79–3.97 (m, 8/3H), 3.96 (dd, 1H, J=4.5, 7.9
Hz), 4.21 (dd, 2/3H, J=4.1, 8.2 Hz), 4.30* (dd, 1/3H,
J=4.1, 8.2 Hz), 4.41–4.51 (m, 3H), 4.54 and 4.57 (AB
type d’s, each 1H, J=11.7 Hz), 4.81* (dd, 1/3H, J=3.4,
9.3 Hz), 4.85* (d, 1/3H, J=10.7 Hz), 5.24–5.29 (m, 2/
3H), 6.24 (d, 2/3H, J=8.4 Hz), 6.53* (d, 1/3H, J=8.4
Hz), 6.70 (br s, 1H), 6.89* (d, 1/3H, J=9.4 Hz), 7.13*
(dd, 1/3H, J=2.9, 6.8 Hz), 7.27–7.39 (m, 5H), 7.63 (dd,
2/3H, J=3.9, 8.8 Hz), 8.14 (br s, 2/3H); 13C NMR
(125 MHz, CDCl3, both rotamers) d ppm: 10.0, 10.2,
10.80, 10.84, 14.0, 14.9, 15.0, 15.1, 21.7, 22.5, 22.57,
22.60, 22.7, 23.1, 23.2, 23.9, 24.0, 24.1, 24.9, 25.2, 26.4,
27.0, 27.1, 29.0, 29.2, 29.4, 31.5, 31.6, 31.7, 37.4, 37.7,
38.1, 38.5, 39.7, 40.5, 40.7, 54.6, 54.7, 55.8, 56.2, 56.3,
56.5, 59.3, 67.8, 68.1, 73.5, 73.8, 76.3, 77.5, 78.2, 127.9,
128.1, 128.3, 128.5, 128.7, 128.8, 136.6, 137.0, 168.9,
169.5, 169.90, 169.94, 171.1, 171.9, 172.8, 173.1, 174.2,
174.3, 177.1; IR (CHCl3) cmÀ1: 3429, 3360, 2963, 2932,
2873, 1667, 1543, 1503, 1467, 1101; HRMS calcd for
C39H64N5O8 (M+H)+ 730.4755, found 730.4752.
4.1.62. 1-N-Demethyl-24-OBn-globomycin (22j). The
reaction was carried out in THF as a solvent. 42% (two
steps), a colorless solid (mp 197–199 ꢀC), 1H NMR
(500 MHz, CD3OD) d ppm: 0.88–0.96 (m, 15H), 1.16 (d,
3H, J=7.3 Hz), 1.21 (d, 3H, J=6.5 Hz), 1.25–1.31 (m,
9H), 1.46–1.53 (m, 2H), 1.63–1.74 (m, 4H), 2.00–2.08
(m, 1H), 2.61 (qu, 1H, J=7.5 Hz), 3.68 (d, 1H, J=17.2
Hz), 3.79 (dd, 1H, J=4.4, 9.9 Hz), 3.95 (dd, 1H, J=5.3,
9.9 Hz), 4.11–4.13 (m, 1H), 4.20 (t, 1H, J=4.8 Hz),
4.26–4.29 (m, 1H), 4.29 (d, 1H, J=4.2 Hz), 4.38 (d, 1H,
J=17.2 Hz), 4.40 (d, 1H, J=4.9 Hz), 4.54 and 4.55 (AB
type d’s, each 1H, J=11.9 Hz), 5.21 (dt, 1H, J=3.9, 8.1
Hz), 7.27–7.35 (m, 5H); 13C NMR (100 MHz, CD3OD)
d ppm: 12.1, 14.5, 15.5, 15.6, 19.5, 21.4, 23.70, 23.73,
26.07, 26.12, 27.5, 30.2, 32.8, 33.5, 37.9, 41.9, 42.0, 47.4,
54.7, 57.9, 59.2, 60.5, 68.5, 69.0, 74.3, 76.7, 128.7, 128.9,
129.3, 138.7, 170.8, 171.5, 172.2, 174.8, 175.7 176.7; IR
(KBr) cmÀ1: 3322, 2959, 2930, 2873, 1739, 1664, 1525,
1456, 1210, 1101; HRMS calcd for C38H62N5O9
(M+H)+ calcd 732.4548, found 732.4561. Anal. calcd
.
for C38H61N5O9 1/2H2O: C, 61.60; H, 8.43; N, 9.45;
Found: C, 61.42; H, 8.59; N, 9.21; [a]2D4 +9.9 (c 0.54,
CH3OH).
.
Anal. calcd for C39H63N5O8 H2O: C, 62.63; H, 8.76; N,
9.36, found: C, 62.66; H, 8.44; N, 9.26; [a]2D4 +15.3 (c
2.86, CHCl3).
4.1.63. 1-N-Et derivative (22k). 54% (two steps), a col-
orless solid (mp, 78 ꢀC), H NMR [500 MHz, CDCl3,
1
major conformer (major/minor>20/1)] d ppm: 0.88 (t,
3H, J=6.8 Hz), 0.92–0.98 (m, 12H), 1.12 (d, 3H, J=7.0
Hz), 1.21–1.33 (m, 14H), 1.36–1.55 (m, 4H), 1.59–1.68
(m, 1H), 1.84–2.05 (m, 3H), 2.94–3.01 (m, 1H), 3.41–
3.49 (m, 2H), 3.53 (dd, 1H, J=3.2, 17.0 Hz), 3.56–3.60
(m, 2H), 3.79 (dd, 1H, J=4.3, 10.0 Hz), 3.89 (dd, 1H,
J=5.2, 10.0 Hz), 4.01–4.09 (m, 1H), 4.22 (q, 1H, J=4.1
Hz), 4.32 (dd, 1H, J=3.7, 7.8 Hz), 4.41 (dd, 1H, J=3.7,
7.8 Hz), 4.48 (dd, 1H, J=9.0, 17.0 Hz), 4.52 and 4.56
(AB type d’s, each 1H, J=11.9 Hz), 5.30–5.35 (m, 1H),
6.59 (d, 1H, J=2.7 Hz), 6.66 (d, 1H, J=8.4 Hz), 7.30–
7.39 (m, 5H), 7.77 (dd,1H, J=3.2, 9.0 Hz), 8.43 (d, 1H,
J=7.8 Hz); 13C NMR (125 MHz, CDCl3, major con-
former) d ppm: 11.8, 14.0, 14.56, 14.62, 15.6, 20.0, 22.3,
22.6, 22.8, 24.1, 25.2, 27.2, 29.2, 31.5, 31.6, 37.3, 39.0,
4.1.61. 22-OMe-24-OBn-globomycin (22h). 37% (2
a
colorless amorphous foam, 1H NMR
steps),
[500 MHz, CDCl3, two rotamers (major/minor=2.0/1)]
d ppm: 0.87–0.99 (m, 12H), 1.12 (d, 2H, J=6.8 Hz),
1.15* (d, 1H, J=6.8 Hz), 1.17 (d, 2H, J=6.8 Hz), 1.19*
(d, 1H, J=6.8 Hz), 1.22–1.39 (m, 8H), 1.43–1.57 (m,
2H), 1.60–1.75 (m, 20/3H), 1.88–1.97 (m, 1H), 2.05–
2.09* (m, 1/3H), 2.10–2.16* (m, 1/3H), 2.19–2.24 (m, 2/
3H), 2.77 (s, 1H), 2.99–3.05 (m, 2/3H), 3.10–3.14* (m, 1/
3H), 3.17 (s, 2H), 3.30 (s, 2H), 3.31 (s, 1H), 3.56 (dd, 2/
3H, J=3.9, 17.6 Hz), 3.69 (dd, 2/3H, J=5.9, 18.6 Hz),
3.79–3.83* (m, 1/3H), 3.85–3.89 (m, 4/3H), 3.96* (dd, 1/
3H, J=3.9, 10.8 Hz), 4.01–4.06 (m, 4/3H), 4.23 (dd, 2/

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
357
40.5, 41.5, 47.0, 56.5, 56.6, 58.0, 67.1, 68.05, 68.09, 73.4,
76.6, 127.9, 128.3, 128.7, 137.0, 168.9, 169.2, 170.3,
173.5, 174.8, 177.1; IR (CHCl3) cmÀ1: 3421, 3339, 2963,
2933, 2873, 1754, 1686, 1665, 1545, 1502; HRMS calcd
for C40H66N5O9 (M+H)+ 760.4861, found 760.4832.
Anal. calcd for C40H65N5O9 1/2H2O: C, 62.48; H, 8.65;
N, 9.11, found: C, 62.40; H, 8.91; N, 8.94; [a]2D5 +56.4
(c 1.33, CHCl3).
(m, 9H), 0.92 (d, 3H, J=6.7 Hz), 0.97 (d, 3H, J=6.8
Hz), 1.22 (d, 3H, J=6.4 Hz), 1.26–1.54 (m, 11H), 1.56–
1.73 (m, 3H), 1.88–1.98 (m, 2H), 2.50 (dd, 1H, J=12.0,
13.3 Hz), 2.78 (dd, 1H, J=1.7, 13.3 Hz), 2.94 (s, 3H),
3.11 (br, 1H), 3.69 (dd, 1H, J=2.9, 18.1 Hz), 3.79 (dd,
1H, J=4.4, 9.9 Hz), 3.91 (dd, 1H, J=4.9, 9.9 Hz), 3.98–
4.04 (m, 1H), 4.21–4.24 (m, 1H), 4.47 (dd, 1H, J=6.7,
9.3 Hz), 4.53 and 4.60 (AB type d’s, each 1H, J=11.8
Hz), 4.58–4.61 (m, 1H), 4.87 (dd, 1H, J=3.4, 9.1 Hz),
5.17–5.19 (m, 1H), 5.28–5.30 (m, 1H), 6.52–6.55 (m,
2H), 7.06 (d, 1H, J=9.1 Hz), 7.27–7.40 (m, 5H), 7.55
(dd, 3H, J=2.4, 8.8 Hz); 13C NMR (125 MHz, CDCl3,
both rotamers) d ppm: 11.9, 14.0, 14.7, 15.4, 19.8, 21.4,
21.9, 22.5, 23.3, 23.4, 24.7, 25.2, 27.1, 27.3, 28.9, 29.7,
31.5, 31.6, 32.4, 35.1, 36.4, 38.2, 39.8, 40.4, 56.0, 56.5,
56.6, 67.8, 68.2, 71.4, 73.5, 73.7, 127.97, 128.04, 128.4,
128.7, 136.8, 169.2, 171.1, 171.4, 172.1, 173.7; IR
(CHCl3) cmÀ1: 3428, 3367, 2961, 2931, 2873, 1740, 1666,
1512, 1386, 1251; HRMS calcd for C38H61N5O9Na
(M+Na)+ 754.4367, found 754.4380. Anal. calcd for
.
4.1.64. Proline-substituted derivative (22l). 52% (two
steps), a colorless solid (mp, 99–101 ꢀC), 1H NMR
(500 MHz, CDCl3, major conformer) d ppm: 0.77 (d,
3H, J=6.8 Hz), 0.88 (t, 3H, J=6.8 Hz), 0.94 (t, 3H,
J=7.3 Hz), 1.13 (d, 3H, J=6.8 Hz), 1.23–1.34 (m, 14H),
1.48–1.56 (m, 1H), 2.00–2.12 (m, 3H), 2.18–2.25 (m,
1H), 2.31–2.40 (m, 1H), 2.89 (qu, 1H, J=6.8 Hz), 3.66–
3.73 (m, 3H), 3.82 (dd, 2H, J=4.9, 9.8 Hz), 4.02 (dd,
1H, J=4.9, 16.6 Hz), 3.95 (dd, 1H, J=4.9, 16.6 Hz),
4.33–4.38 (m, 2H), 4.40–4.45 (m, 2H), 4.52 and 4.53
(AB type d’s, each 1H, J=11.7 Hz), 4.82–4.88 (m, 1H),
5.04–5.08 (m, 1H), 5.78 (d, 1H, J=6.8 Hz), 7.26–7.40
(m, 5H), 7.51 (br s, 1H), 7.56 (br d, 1H, J=6.8 Hz), 7.69
(br d, 1H, J=6.8 Hz); 13C NMR (125 MHz, CDCl3,
both rotamers) d ppm: 11.7, 11.8, 13.67, 13.72, 14.0,
14.3, 15.4, 19.6, 22.5, 22.6, 25.1, 25.2, 25.4, 27.0, 29.0,
29.1, 29.4, 29.7, 31.6, 33.0, 35.2, 41.9, 42.1, 48.1, 52.1,
57.9, 59.6, 62.6, 66.8, 69.4, 73.4, 73.6, 75.2, 76.1, 76.4,
127.88, 127.98, 128.03, 128.1, 128.4, 128.5, 137.2, 168.4,
.
C38H61N5O9 1/2H2O: C, 61.60; H, 8.43; N, 9.45, found:
C 61.75; H, 8.10; N, 9.23; [a]2D6 À59.6 (c 1.09, CHCl3).
4.1.67. 5-Aza-24-OBn-globomycine (22o). 61% (two
steps), a colorless sold (mp 88 ꢀC), H NMR (400 MHz,
1
CDCl3) d ppm: 0.86–0.89 (m, 6H), 0.95 (t, 3H, J=7.3
Hz), 1.00–1.14 (m, 9H), 1.18 (d, 3H, J=6.3 Hz), 1.21–
1.43 (m, 10H), 1.58–1.69 (m, 2H), 1.82–2.04 (m, 3H),
2.30–2.37 (m, 1H), 3.01 (br s, 1H), 3.23 (s, 3H), 3.40 (br
s, 2H), 3.48 (dd, 1H, J=4.0, 17.0 Hz), 3.61 (br s, 1H),
3.95 (d, 2H, J=7.0 Hz), 4.24–4.25 (m, 1H), 4.35 (dd,
1H, J=8.7, 17.0 Hz), 4.50–4.51 (m, 1H), 4.55 and 4.59
(AB type d’s, each 1H, J=11.8 Hz), 4.71 (q, 1H, J=7.0
Hz), 4.77 (br s, 1H), 5.95 (d, 1H, J=7.0 Hz), 6.70 (br s,
1H), 7.07 (br d, 1H, J=5.8 Hz), 7.29–7.36 (m, 5H), 8.12
(br s, 1H), 8.23 (br d, 1H, J=7.0 Hz), 13C NMR
(125 MHz, CDCl3) d ppm: 11.8, 14.1, 14.6, 16.0, 19.4,
22.3, 22.7, 23.2, 25.4, 26.3, 26.9, 28.8, 29.7, 31.7, 35.1,
37.9, 38.8, 43.1, 52.0, 57.3, 59.1, 63.4, 65.7, 67.1, 73.4,
127.5, 127.7, 128.3, 137.9, 168.8, 171.2, 171.7, 171.9,
173.3, 176.4; IR (CHCl3) cmÀ1: 3374, 3302, 2962, 2932,
2873, 1665, 1621, 1544, 1287, 1110; HRMS calcd for
C39H64N6O8Na (M+Na)+ 767.4683, found 767.4683.
170.4, 170.7, 171.5, 172.0, 172.3, 174.6; IR (KBr) cmÀ1
:
3338, 2961, 2931, 2873, 1741, 1658, 1525, 1456, 1261,
1099; HRMS calcd for C37H57N5O9Na (M+Na)+
calcd 738.4054, found 738.4055. Anal. calcd for
.
C37H57N5O9 1/2H2O: C, 61.31; H, 8.06; N, 9.66, found:
C, 61.13; H, 7.93; N, 9.64; [a]2D7 À34.3 (c 1.14, CHCl3).
4.1.65. Homo-proline-substituted derivative (22m). 16%
(two steps), a colorless solid (mp, 86 ꢀC), 1H NMR
(400 MHz, CDCl3, major conformer) d ppm: 0.86–0.92
(m, 6H), 0.91 (d, 3H, J=6.8 Hz), 0.98 (t, 3H, J=6.8
Hz), 1.14 (d, 3H, J=6.8 Hz), 1.21 (d, 3H, J=5.9 Hz),
1.24–1.45 (m, 11H), 1.48–1.67 (br m, 2H), 1.67–1.75 (br
m, 1H), 1.82–1.92 (br m, 1H), 1.97–2.05 (br m, 1H),
2.45–2.53 (br m, 2H), 2.94–3.02 (br m, 1H), 3.79–3.87
(m, 2H), 3.94 (br s, 1H), 3.98–4.08 (br m, 1H), 4.16 (dd,
1H, J=7.3, 18.1 Hz), 4.27–4.33 (m, 2H), 4.45–4.67 (m,
4H), 4.75 (dd, 1H, J=3.9, 8.8 Hz), 5.09 (br d, 1H,
J=8.8 Hz), 6.53 (d, 1H, J=8.8 Hz), 6.83 (br s, 1H), 7.08
(d, 1H, J=8.8 Hz), 7.28–7.37 (m, 5H), 7.46 (br s, 1H);
13C NMR (125 MHz, CDCl3, both rotamers) d ppm:
11.9, 13.0, 14.2, 15.0, 19.9, 21.2, 22.7, 25.0, 25.6, 26.9,
27.1, 29.2, 29.8, 30.0, 31.7, 31.8, 37.8, 39.7, 40.3, 41.2,
54.8, 56.2, 57.2, 57.4, 67.7, 67.9, 73.5, 73.7, 76.8, 76.9,
127.6, 128.0, 128.3, 128.4, 128.6,136.4, 169.4, 169.5,
169.8, 171.1, 172.5, 173.7; IR (CHCl3) cmÀ1: 3690, 3422,
3361, 2931, 2861, 1737, 1676, 1503, 1425, 1429; HRMS
calcd for C38H59N5O9Na (M+Na)+ 752.4210, found
.
Anal. calcd for C39H64N6O8 H2O: C, 61.39; H, 8.72; N,
11.01, found: C 61.48; H, 8.50; N, 10.77; [a]2D4 À153.5 (c
1.30, CHCl3).
4.1.68. 4-epi-24-OBn-Globomycin (22p). 39% (two
steps), a colorless solid (mp 82 ꢀC), 1H NMR [400 MHz,
CDCl3, two rotamers (major/minor=1.4/1)] d ppm:
0.73* (d, 6/5H, J=6.8 Hz), 0.82–0.95 (m, 51/5H), 0.97
(d, 9/5H, J=6.5 Hz), 1.00 (d, 9/5H, J=6.4 Hz), 1.13–
1.32 (m, 16H), 1.39–1.45 (m, 7/5H), 1.57–1.66 (m, 2H),
1.79–1.86 (m, 3/5H), 1.96–2.03 (m, 3/5H), 2.18–2.20*
(m, 2/5H), 2.35–2.37* (m, 2/5H), 2.49–2.52 (m, 3/5H),
2.84 (s, 6/5H), 2.89–2.92 (m, 3/5H), 3.06–3.10* (m, 2/
5H), 3.19 (s, 9/5H), 3.22* (d, 2/5H, J=9.4 Hz), 3.28 (d,
3/5H, J=8.7 Hz), 3.61 (dd, 1H, J=5.3, 9.6 Hz), 3.67
(dd, 1H, J=4.9, 9.6 Hz), 3.85–3.96 (m, 13/5H), 4.15*
(dd, 2/5H, J=4.1, 7.5 Hz), 4.18–4.28 (m, 8/5H), 4.33–
4.46 (m, 8/5H), 4.51 and 4.72* (AB type d’s, each 2/5H,
J=11.8 Hz), 4.53–4.60 (m, 8/5H), 5.01* (br s, 2/5H),
5.16–5.20 (m, 3/5H), 5.21–5.26* (m, 2/5H), 6.30* (d, 2/
.
752.4221. Anal. calcd for C38H59N5O9 2/3H2O: C,
61.52; H, 8.20; N, 9.44, found: C, 61.48; H, 8.07; N,
9.32; [a]2D6 À42.6 (c 1.86, CHCl3).
4.1.66. 3-Demethyl-24-OBn-globomycin (22n). 55% (2
a
colorless amorphous foam, 1H NMR
steps),
(400 MHz, CDCl3, major conformer) d ppm: 0.86–1.02

358
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
5H, J=8.7 Hz), 6.84–6.86* (m, 2/5H), 6.94–6.95 (m, 3/
5H), 7.28–7.38 (m, 33/5H), 7.89* (d, 2/5H, J=9.6 Hz),
8.36 (br s, 3/5H); 13C NMR (125 MHz, CDCl3, both
rotamers) d ppm: 11.4, 11.7, 14.0, 14.5, 15.3, 15.7, 19.5,
20.3, 21.9, 22.4, 22.5, 22.6, 23.1, 25.0, 25.3, 25.8, 25.9,
26.4, 27.0, 28.92, 28.94, 29.7, 31.3, 31.6, 31.7, 33.7, 34.6,
35.2, 37.59, 37.64, 39.5, 40.0, 41.3, 41.5, 41.9, 51.4, 54.9,
56.6, 56.8, 57.8, 60.1, 66.4, 67.6, 67.8, 68.1, 68.3, 73.6,
74.1, 77.9, 127.8, 127.9, 128.2, 128.4, 128.5, 128.7, 136.4,
137.2, 167.2, 169.6, 170.2, 170.4, 170.5, 170.6, 172.4,
172.9, 174.6, 175.7; IR (CHCl3) cmÀ1: 3394, 2963, 2931,
2873, 1727, 1663, 1535, 1498, 1261, 1100; HRMS calcd
for C39H63N5O9Na (M+Na)+ 768.4523; Found
Hz), 1.09 (d, 9/4H, J=6.9 Hz), 1.14* (d, 3/4H, J=6.9
Hz), 1.16–1.47 (m, 13H), 1.49–1.73 (m, 15/4H), 1.82–
1.87* (m, 1/4H), 1.92–2.07* (m, 2/4H), 2.10–2.16 (m, 6/
4H), 2.79* (s, 3/4H), 2.86 (br, 2H), 3.12–3.19 (m, 1H),
3.21 (s, 9/4H), 3.63 (br s, 3/4H), 3.81 (dd, 3/4H, J=3.8,
17.3 Hz), 3.87* (dd, 1/4H, J=3.7, 18.0 Hz), 3.93–4.03
(m, 10/4H), 4.08 (t, 3/4H, J=6.6 Hz), 4.17–4.20* (m, 1/
4H), 4.24 (dd, 3/4H, J=7.2, 17.3 Hz), 4.30 (d, 3/4H,
J=6.8 Hz), 4.36–4.41 (m, 7/4H), 4.52* (t, 1/4H, J=7.0
Hz), 4.59* (dd, 1/4H, J=5.8, 9.1 Hz), 4.95* (d, 1/4H,
J=9.3 Hz), 5.03 (dt, 3/4H, J=3.1, 8.6 Hz), 6.90* (d, 1/
4H, J=9.1 Hz), 7.18 (d, 3/4H, J=7.5 Hz), 7.35 (br s, 3/
4H), 7.51–7.53* (br, 2/4H), 7.65–7.68 (br m, 7/4H); 13C
NMR (100 MHz, CDCl3, 33 mM, both rotamers) d
ppm: 11.6, 12.0, 12.7, 14.2, 15.1, 16.0, 16.5, 18.9, 19.1,
22.0, 22.7, 22.9, 23.1, 23.2, 24.5, 24.9, 25.0, 25.4, 25.7,
29.18, 29.23, 29.5, 29.8, 31.5, 31.7, 31.8, 36.5, 37.4, 38.1,
38.6, 39.0, 40.2, 40.8, 41.0, 41.5, 57.1, 57.5, 57.6, 57.8,
58.6, 59.0, 59.1, 60.7, 61.6, 66.7, 67.1, 76.5, 76.9, 77.7,
168.7, 170.38, 170.44, 170.58, 170.66, 170.9, 172.7,
173.7, 174.4, 176.4; IR (KBr) cmÀ1: 3327, 2960, 2931,
2873, 1739, 1656, 1543, 1466, 1378, 1199; HRMS calcd
for C32H58N5O9 (M+H)+ calcd 656.4235, found
.
768.4547. Anal. calcd for C39H63N5O9 1/3H2O: C,
62.29; H, 8.53; N, 9.31, found: C 62.18; H, 8.24; N, 8.91;
[a]2D4 +15.4 (c 1.65, CHCl3).
4.1.69. 8-N-Me-24-OBn-globomycin (22q). 41% (2
steps), a colorless solid (mp 63–65 ꢀC), 1H NMR
(500 MHz, CDCl3, major conformer) d ppm: 0.83 (d,
3H, J=6.8 Hz), 0.86–0.98 (m, 12H), 1.18 (d, 3H, J=6.8
Hz), 1.20 (d, 3H, J=6.8 Hz), 1.23–1.46 (m, 7H), 1.47–
1.59 (m, 2H), 1.63–1.79 (m, 4H), 1.81–1.99 (m, 2H),
2.15–2.27 (m, 2H), 3.04 (s, 3H), 3.17–3.21 (m, 1H), 3.27
(s, 3H), 3.59 (d, 1H, J=8.8 Hz), 3.62 (d, 1H, J=8.8
Hz), 3.86 (dd, 1H, J=5.9, 8.8 Hz), 4.05–4.09 (m, 1H),
4.33 (br d, 1H, J=6.8 Hz), 4.38–4.42 (m, 1H), 4.45 (dd,
1H, J=2.9, 8.8 Hz), 4.53 (d, 1H, J=11.2 Hz), 4.56 (d,
1H, J=8.8 Hz), 4.66 (d, 1H, J=11.2 Hz), 4.76–4.81 (m,
1H), 5.11–5.15 (m, 1H), 7.11 (br d, 1H, J=4.8 Hz),
7.27–7.42 (m, 6H), 8.03 (br s, 1H); 13C NMR (125 MHz,
CDCl3, both rotamers) d ppm: 10.9, 11.7, 11.9, 12.3,
14.0, 14.1, 15.3, 19.47, 19.50, 21.9, 22.51, 22.55, 22.59,
22.9, 23.0, 24.3, 25.3, 25.5, 26.3, 26.9, 27.4, 28.1, 28.9,
29.0, 30.9, 31.6, 31.8, 35.5, 36.0, 37.0, 37.2, 38.2, 38.3,
38.5, 40.6, 41.7, 50.0, 51.9, 52.2, 54.0, 55.7, 56.4, 59.0,
68.4, 68.6, 68.8, 69.2, 73.4, 76.3, 78.5, 127.7, 127.8,
128.1, 128.19, 128.25, 128.33, 128.6, 137.5, 168.0,
169.92, 169.99, 170.01, 170.2, 170.8, 171.4, 172.2, 172.6,
177.5; IR (KBr) cmÀ1: 3380, 2958, 2930, 2873, 1745,
1681, 1657, 1530, 1496, 1204; HRMS calcd for
C40H65N5O9Na (M+Na)+ calcd 782.4680, found
.
656.4227. Anal. calcd for C32H57N5O9 3/4H2O: C,
57.42; H, 8.81; N, 10.46, found: C, 57.44; H, 8.67; N,
10.27; [a]2D4 +3.1 (c 0.50, CH3OH).
4.2.2. 22-epi-ꢀGlobomycin (1e). 90%, a colorless solid
1
(mp110–112 C), H NMR [500 MHz, CDCl3, 16 mM,
major conformer (major/minor=8.8/1)] d ppm: 0.87–
0.97 (m, 15H), 1.12 (d, 3H, J=6.8 Hz), 1.20 (d, 3H,
J=5.9 Hz), 1.30–1.42 (m, 10H), 1.44–1.57 (m, 2H),
1.60–1.68 (m, 1H), 1.69–1.74 (m, 1H), 2.13–2.21 (m,
2H), 2.30 (br s, 2H), 3.03–3.09 (m, 1H), 3.22 (s, 3H),
3.62 (s, 1H), 3.65 (s, 1H), 3.86 (dd, 1H, J=5.9, 11.7 Hz),
4.00 (dd, 1H, J=3.9, 11.7 Hz), 4.18 (s, 1H), 4.35 (d, 1H,
J=8.8 Hz), 4.42–4.45 (m, 1H), 4.64 (dd, 1H, J=3.9, 7.8
Hz), 4.69 (q, 1H, J=5.9 Hz), 5.31–5.35 (m, 1H), 6.74 (d,
1H, J=8.8 Hz), 7.55 (s, 1H), 8.02 (d, 1H, J=3.9 Hz),
8.43 (br s, 1H); 13C NMR (100 MHz, CDCl3, both
rotamers) d ppm: 11.76, 11.85, 13.0, 14.2, 14.5, 14.9,
15.1, 19.6, 19.8, 22.0, 22.7, 23.1, 23.3, 24.1, 25.2, 25.3,
25.6, 26,9, 27.4, 29.3, 29.8, 31.6, 31.7, 31.8, 37.2, 37.7,
38.4, 38.9, 39.4, 40.3, 40.8, 41.6, 56.6, 57.8, 58.9, 59.1,
60.9, 61.8, 65.9, 66.3, 69.1, 76.5, 77.6, 169.3, 169.7,
170.50, 170.56, 170.62, 170.9, 173.3, 173.6, 174.8, 175.2,
177.1; IR (KBr) cmÀ1: 3330, 2959, 2931, 2873, 1740,
1657, 1543, 1467, 1379, 1204; HRMS calcd for
C32H58N5O9 (M+H)+ calcd 656.4235, found 656.4232;
[a]2D4 +34.4 (c 0.50, CH3OH).
.
782.4671. Anal. calcd for C40H65N5O9 1/5H2O: C,
62.92; H, 8.63; N, 9.17, found: C, 62.51; H, 8.01; N,
9.03; [a]2D6 À21.6 (c 1.01, CHCl3).
4.2. Globomycin analogues
Compound 22c (28.2 mg, 35.2 mmol) was dissolved in
CH3OH (1.5 mL). To this solution, Pd(OH)2 (23.6 mg)
was added and the resulting mixture was stirred at room
temperature for 11 h under H2 atmosphere. Pd(OH)2
was removed by filtration and the filtrate was evapo-
rated. The residue was purified by column chromato-
graphy with a gradient elution system, 30:1–20:1
mixture of CH2Cl2 and CH3OH as eluents to give 1c1f
(22.6 mg, 31.7 mmol, 90%) as a colorless solid (mp92–
94 ꢀC).
4.2.3. Stereoisomer at the C(22) and C(25) positions of
globomycin (1f). 95%, a colorless solid (mp, 118–
120 ꢀC), 1H NMR [500 MHz, CDCl3, 16 mM, two
rotamers (major/minor=5.2/1)] d ppm: 0.88 (t, 3H,
J=6.9 Hz), 0.92–0.97 (m, 9H), 1.03 (d, 3H, J=6.7 Hz),
1.10 (d, 3H, J=6.9 Hz), 1.19 (d, 3H, J=6.5 Hz), 1.20–
1.33 (m, 10H), 1.34–1.42 (m, 1H), 1.46–1.60 (m, 1H),
1.60 (br s, 2H), 1.66–1.73 (m, 1H), 1.92–1.98* (m, 1/
6H), 2.04–2.11 (m, 1H), 2.16–2.22 (m, 5/6H), 2.79* (s, 3/
6H), 2.98 (br s, 1H), 3.05 (qu, 5/6H, J=7.3 Hz), 3.16–
3.21* (m, 1/6H), 3.22 (s, 15/6H), 3.48* (d, 1/6H, J=4.1
Hz), 3.64 (br s, 5/6H), 3.81 (dd, 5/6H, J=7.8, 18.0 Hz),
4.2.1. 25-epi-Globomycin (1d). 97%, a colorless solid
(mp108–110 ꢀC), H NMR [500 MHz, CDCl3, 16 mM,
1
two rotamers (major/minor=4.5/1)] d ppm: 0.88 (t, 3H,
J=6.7 Hz), 0.90–1.05 (m, 39/4H), 1.01 (d, 9/4H, J=6.8

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
359
3.83–3.93 (m, 1H), 3.97–4.13 (m, 9/6H), 4.24–4.35 (m,
15/6H), 4.39* (d, 1/6H, J=8.1 Hz), 4.48 (dd, 5/6H,
J=5.2, 8.0 Hz), 4.52* (t, 1/6H, J=6.3, 9.3 Hz), 4.59*
(dd, 1/6H, J=6.3, 9.3 Hz), 4.62–4.68 (m, 5/6H), 5.05*
(d, 1/6H, J=9.5 Hz), 5.26–5.30 (m, 5/6H), 6.75 (d, 5/
6H, J=8.6 Hz), 6.78* (m, 1/6H), 7.41 (d, 5/6H, J=3.1
Hz), 7.57* (d, 1/6H, J=8.4 Hz), 7.62* (br s, 1/6H),
7.78* (br t, 1/6H, J=2.4 Hz), 7.95 (br s, 5/6H), 8.16 (br
s, 5/6H); 13C NMR (125 MHz, CDCl3, both rotamers) d
ppm: 10.9, 11.8, 13.4, 14.02, 14.05, 14.09, 15.0, 15.6,
16.3, 19.6, 19.7, 21.8, 22.4, 22.56, 22.62, 23.0, 23.2, 24.1,
24.2, 24.5, 24.8, 25.3, 25.5, 29.1, 29.2, 29.7, 29.9, 31.5,
31.56, 31.60, 31.7, 36.7, 37.4, 38.2, 38.6, 39.2, 40.2, 40.8,
41.2, 42.2, 56.9, 57.1, 58.0, 58.4, 58.6, 58.9, 60.6, 61.9,
65.7, 66.3, 68.4, 76.4, 169.4, 169.9, 170.2, 170.85, 170.9,
171.2, 171.4, 172.7, 173.4, 174.4, 175.1, 177.2; IR (KBr)
cmÀ1: 3327, 2959, 2931, 2873, 1738, 1656, 1543, 1520,
1466, 1203; HRMS calcd for C32H57N5O9Na
(M+Na)+ calcd 678.4054, found 678.4044. Anal. calcd
(dd, 1/5H, J=3.9, 7.8 Hz), 4.80 (dd, 1/5H, J=3.9, 8.8
Hz), 4.83 (dd, 4/5H, J=3.9, 8.8 Hz), 4.94 (d, 1/5H,
J=10.7 Hz), 5.27–5.31 (m, 1H), 6.46 (d, 4/5H, J=7.8
Hz), 6.89* (d, 1/5H, J=9.8 Hz), 7.06* (d, 1/5H, J=7.8
Hz), 7.28-7.30 (m, 4/5H), 7.47 (d, 1H, J=3.9 Hz), 7.84–
7.95 (br s, 1H), 8.09 (br s, 1H); 13C NMR (125 MHz,
CHCl3, both rotamers) d ppm: 11.8, 12.6, 14.2, 14.9,
15.0, 15.1, 15.2, 21.9, 22.7, 23.2, 23.3, 24.3, 25.2, 25.3,
25.9, 27.0, 27.3, 29.3, 29.6, 29.7, 29.8, 31.6, 31.7, 31.8,
37.0, 37.5, 38.3, 38.7, 39.4, 40.3, 41.3, 41.6, 55.1, 55.7,
56.3, 56.4, 56.57, 56.63, 57.8, 58.6, 59.3, 61.1, 61.8, 68.5,
74.9, 75.3, 76.0, 76.5, 77.2, 77.7, 168.5, 169.0, 169.1,
169.9, 170.2, 170.7, 173.4, 174.7, 174.9, 177.1; IR
(CHCl3) cmÀ1: 3347, 2962, 2932, 1668, 1546, 1501,
1467, 1380, 1249, 1187; HRMS calcd for C33H60N5O9
(M+H)+ 670.4391, found 670.4403. Anal. calcd for
.
C33H59N5O9 H2O: C, 57.62; H, 8.94; N, 10.18, found:
C 57.91; H, 8.94; N, 10.03; [a]2D5 +20.9 (c 2.69,
CHCl3).
.
for C32H57N5O9 H2O: C, 57.04; H, 8.83; N, 10.39,
found: C, 57.23; H, 8.54; N, 10.32; [a]2D7 +16.1 (c 0.50,
CH3OH).
4.2.6. 24-OMe-Globomycin (1i). 27%, a colorless amor-
1
phous foam, H NMR [500 MHz, CDCl3, two rotamers
(major/minor=3.7/1)] d ppm: 0.82–0.99 (m, 14H), 1.12
(d, 12/5H, J=6.8 Hz), 1.17* (d, 3/5H, J=6.8 Hz), 1.33
(d, 3H, J=6.8 Hz), 1.52–2.00 (m, 15H), 2.00–2.09 (m,
1H), 2.10–2.17 (m, 1H), 2.79* (s, 3/5H), 3.07–3.16 (m,
1H), 3.19 (s, 12/5H), 3.39 (s, 12/5H), 3.42* (s, 3/5H),
3.61 (dd, 4/5H, J=3.9, 17.6 Hz), 3.70 (br s, 1H), 3.67–
3.83 (m, 16/5H), 4.02–4.04* (m, 1/5H), 4.10–4.15 (m, 4/
5H), 4.25–4.32 (m, 8/5H), 4.43 and 4.46 (AB type d’s,
each 1H, J=8.8 Hz), 4.50–4.53* (m, 1/5H), 4.80 (dd,
1H, J=3.9, 8.8 Hz), 4.90 (d, 1H, J=10.7 Hz), 5.16–
5.20* (m, 1/5H), 6.70* (br d, 1/5H, J=3.9 Hz), 6.81–
6.85 (m, 9/5H), 7.18* (d, 1/5H, J=7.8 Hz), 7.34* (d, 1/
5H, J=8.8 Hz), 7.69 (dd, 4/5H, J=2.9, 7.8 Hz), 8.00 (br
s, 4/5H); 13C NMR (125 MHz, CDCl3, both rotamers) d
ppm: 11.9, 14.2, 14.8, 15.2, 20.0, 20.1, 21.9, 22.7, 23.2,
23.3, 24.3, 24.9, 25.3, 26.3, 27.1, 27.3, 28.4, 29.1, 29.3,
29.6, 29.8, 31.4, 31.7, 31.8, 37.0, 37.7, 38.2, 38.5, 39.7,
40.8, 41.5, 45.3, 55.5, 55.7, 56.3, 56.6, 56.8, 57.9, 59.1,
59.3, 67.9, 68.0, 70.1, 70.4, 76.6, 76.9, 77.2, 78.2, 94.4,
123.9, 129.3, 168.96, 169.00, 169.8, 170.0, 170.5, 171.0,
4.2.4. 22-Deoxyglobomycin (1g). 93%, a colorless solid
(mp, 95 ꢀC), H NMR [400 MHz, CDCl3, two rotamers
1
(major/minor=4.0/1)] d ppm: 0.77–1.07 (m, 14H), 0.88
(t, 3H, J=6.5 Hz), 1.11 (d, 12/5H, J=6.9 Hz), 1.15* (d,
3/5H, J=6.9 Hz), 1.26–1.55 (m, 15H), 1.67–1.87 (m,
2H), 1.98–2.07 (m, 1H), 2.09–2.20 (m, 1H), 2.78* (s, 3/
5H), 3.02–3.10 (m, 4/5H), 3.15–3.20* (s, 1/5H), 3.21 (s,
12/5H), 3.55–3.72 (m, 4H), 3.86 (dd, 1H, J=5.9, 11.5
Hz), 3.90–3.91* (m, 1/5H), 3.96 (dd, 4/5H, J=3.9, 11.5
Hz), 4.05–4.07* (m, 1/5H), 4.19–4.24 (m, 4/5H), 4.27–
4.37 (m, 9/5H), 4.52* (t, 1/5H, J=6.9 Hz), 4.66 (dd, 4/
5H, J=3.7, 7.7 Hz), 4.75* (dd, 1/5H, J=4.6, 9.3 Hz),
4.96* (br d, 1/5H, J=10.0 Hz), 5.24–5.29 (m, 4/5H),
6.46 (d, 4/5H, J=7.8 Hz), 6.86* (d, 1/5H, J=9.3 Hz),
7.06* (d, 1/5H, J=7.8 Hz), 7.53–7.55* (m, 1/5H), 7.65*
(br s, 1/5H), 7.78 (d, 4/5H, J=2.9 Hz), 7.80–7.85 (br, 4/
5H), 8.13–8.20 (br, 4/5H); 13C NMR (125 MHz, CDCl3,
both rotamers) d ppm: 10.2, 10.3, 11.60, 11.63, 12.3,
14.0, 14.7, 14.8, 14.9, 21.8, 22.5, 23.0, 23.1, 23.7, 23.9,
24.3, 25.0, 25.2, 25.7, 26.8, 27.1, 29.06, 29.11, 29.2, 29.6,
29.7, 31.5, 31.6, 31.7, 36.8, 37.4, 38.1, 38.6, 39.2, 40.5,
41.3, 41.5, 54.9, 55.1, 56.5, 57.5, 57.8,59.2, 60.8, 61.9,
76.1, 77.7, 168.9, 170.0, 170.7, 170.9, 171.3, 171.4, 173.3,
174.6, 177.2; IR (CHCl3) cmÀ1: 3355, 2963, 2932, 2874,
1733, 1671, 1545, 1503, 1465, 1379; HRMS calcd for
C32H57N5O8Na (M+Na)+ 662.4105, found 662.4114;
[a]2D6 À2.8 (c 2.05, CHCl3).
171.1, 172.4, 172.6, 173.9, 174.2, 176.6; IR (CHCl3) cmÀ1
:
3422, 3342, 2962, 2932, 1737, 1666, 1542, 1511, 1118;
HRMS calcd for C33H59N5O9Na (M+Na)+ 692.4210,
.
found 692.4210. Anal. calcd for C33H59N5O9 9/10H2O:
C, 57.77; H, 8.76; N, 9.84, found: C, 58.06; H, 8.76; N,
9.84; [a]2D5=+13.7 (c 1.19, CHCl3).
4.2.7. 1-N-Demethylglobomycin (1j). 100%, a colorless
solid (mp122–124 ꢀC), H NMR (500 MHz, CD3OD,
1
4.2.5. 22-OMe-globomycin (1h). 99%, a colorless solid
(mp, 89 ꢀC), H NMR [500 MHz, CDCl3, 16 mM, two
single isomer) d ppm: 0.89 (t, 3H, J=7.1 Hz), 0.93–0.99
(m, 9H), 1.00 (d, 3H, J=7.0 Hz), 1.18 (d, 3H, J=7.3
Hz), 1.23 (d, 3H, J=6.5 Hz), 1.25–1.35 (m, 9H), 1.48–
1.58 (m, 2H), 1.64–1.78 (m, 4H), 2.07–2.11 (m, 1H), 2.61
(qu, 1H, J=7.5 Hz), 3.68 (d, 1H, J=17.2 Hz), 3.87 (dd,
1H, J=4.5, 11.2 Hz), 3.97 (dd, 1H, J=4.6, 11.2 Hz),
4.07 (t, 1H, J=4.5 Hz), 4.18–4.21 (m, 1H), 4.28 (dd, 1H,
J=3.9, 10.9 Hz), 4.33 (d, 1H, J=4.1 Hz), 4.39 (d, 1H,
J=17.2 Hz), 4.44 (d, 1H, J=4.6 Hz), 5.24 (dt, 1H,
J=4.5, 8.3 Hz); 13C NMR (125 MHz, CD3OD) d ppm:
12.0, 14.4, 15.5, 19.3, 21.4, 23.61, 23.62, 26.0, 26.1, 27.6,
30.1, 32.8, 33.3, 37.8, 41.9, 42.0, 47.6, 48.3, 54.8, 59.3,
1
rotamers (major/minor=4.3/1)] d ppm: 0.88 (d, 3H,
J=6.8 Hz), 0.90–0.97 (m, 9H), 1.11 (d, 3H, J=6.8 Hz),
1.16* (d, 3/5H, J=6.8 Hz), 1.19 (d, 12/5H, J=6.8 Hz),
1.28–1.38 (m, 12H), 1.41–1.52 (m, 2H), 1.55–1.63 (m,
2H), 1.64–1.72 (m, 1H), 2.07–2.12 (m, 1H), 2.16–2.21
(m, 1H), 2.78* (s, 3/5H), 3.01–3.08 (m, 1H), 3.21 (s, 12/
5H), 3.34 (s, 12/5H), 3.35* (s, 3/5H), 3.62–3.77 (m,
9/5H), 3.90–4.01 (m, 9/5H), 4.05–4.10 (m, 1H), 4.17–
4.25 (m, 1H), 4.30 (dd, 1H, J=7.8, 16.6 Hz), 4.49* (t, 1/
5H, J=6.8 Hz), 4.62 (dd, 4/5H, J=3.9, 7.8 Hz), 4.74*

360
T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
59.7, 60.5, 61.6, 68.4, 76.8, 171.2, 171.8, 173.0, 175.3
176.3, 177.1; IR (KBr) cmÀ1: 3305, 2960, 2931, 2873,
1719, 1656, 1528, 1465, 1381, 1210; HRMS calcd for
C31H55N5O9Na (M+Na)+ calcd 664.3898, found
664.3902. Anal. calcd for C31H55N5O9 11/10H2O: C,
56.28; H, 8.71; N, 10.59, found: C, 56.64; H, 8.46; N,
10.19; [a]2D6 +6.8 (c 0.50, CH3OH).
4.2.10. Homo-proline substituted globomycin (1m). 89%,
a colorless solid (mp104 ꢀC), 1H NMR (400 MHz,
CDCl3, 16 mM, major conformer) d ppm: 0.81–0.99 (m,
12H), 1.14 (d, 3H, J=6.8 Hz), 1.20–1.54 (m, 12H), 1.23
(d, 3H, J=5.9 Hz), 1.55–1.83 (br m, 3H), 2.02–2.12 (br
m, 1H), 2.43–2.56 (br m, 2H), 3.03–3.09 (m, 1H), 3.58–
4.54 (br m, 8H), 4.58–4.72 (br m, 2H), 4.77–4.83 (br m,
1H), 5.07–5.21 (br m, 1H), 6.61 (d, 1H, J=8.8 Hz), 7.41
(br d, 1H, J=5.9 Hz), 7.66 (br s, 1H), 7.71 (br s, 1H);
13C NMR (125 MHz, CDCl3, 29 mM, both rotamers) d
ppm: 11.7, 11.9, 13.3, 14.2, 14.3, 14.8, 15.0, 18.9, 21.2,
22.7, 22.8, 24.9, 25.3, 26.9, 27.1, 29.1, 29.2, 29.4, 29.8,
30.1, 31.7, 31.8, 31.9, 32.0, 37.5, 39.7, 40.2, 41.7, 56.5,
57.4, 58.3, 60.9, 66.9, 76.5, 76.8, 77.2, 169.4, 170.4,
170.7, 170.8, 171.0, 171.1, 171.9, 173.4, 174.1; IR
(CHCl3) cmÀ1: 3690, 3347, 2932, 2861, 1739, 1658, 1512,
1461, 1381, 1249; HRMS calcd for C31H53N5O9Na
(M+Na)+ 662.3741, found 662.3758. Anal. calcd for
.
4.2.8. 1-N-Et-globomycin (1k). 85% a colorless solid
(mp, 86ꢀ), 1H NMR [500 MHz, CDCl3, major con-
former (major/minor >20/1)] d ppm: 0.88 (t, 3H, J=6.8
Hz), 0.90–0.96 (m, 12H), 1.12 (d, 3H, J=6.8 Hz), 1.24–
1.32 (m, 15H), 1.35–1.43 (m, 2H), 1.47–1.57 (m, 2H),
1.88–1.99 (m, 2H), 2.13–2.18 (m, 1H), 2.99 (qu, 1H,
J=6.8 Hz), 3.48 (q, 2H, J=6.8 Hz), 3.56–3.60 (m, 2H),
3.85–3.95 (m, 2H), 3.98–4.04 (br, 2H), 4.14 (d, 1H,
J=2.9 Hz), 4.28–4.33 (m, 2H), 4.44 (dd, 1H, J=9.3,
17.1 Hz), 4.49 (dd, 1H, J=3.4, 7.3 Hz), 5.29 (dt, 1H,
J=2.9, 8.8 Hz), 6.90 (d, 1H, J=6.8 Hz), 7.59 (s, 1H),
7.79 (br d, 1H, J=7.8 Hz), 8.46 (br s, 1H); 13C NMR
(125 MHz, CDCl3, major conformer) d ppm: 11.6, 14.0,
14.4, 14.5, 15.5, 18.5, 22.3, 22.5, 22.7, 24.0, 25.1, 27.2,
29.2, 31.4, 31.5, 37.0, 39.1, 40.1, 41.5, 47.1, 56.5, 58.9,
59.1, 61.0, 66.8, 67.2, 76.2, 168.4, 169.7, 171.1, 174.3,
175.2, 177.3; IR (KBr) cmÀ1: 3325, 2960, 2931, 2873,
1759, 1656, 1547, 1464, 1380, 1193; HRMS calcd for
C33H59N5O9Na (M+Na)+ 692.4210, found 692.4178.
.
C31H53N5O9 H2O: C, 56.60; H, 8.43; N, 10.65; found:
C, 56.89; H, 8.31; N, 10.37; [a]2D6 À56.9 (c 1.93,
CHCl3).
4.2.11. 3-Demethylglobomycin (1n). 94%, a coloreless
powder (mp 107 ꢀC), H NMR [500 MHz, CDCl3, two
1
rotamers (major/minor=4.2/1)] d ppm: 0.87–0.90 (m,
5H), 0.92–1.00 (m, 9H), 1.19 (d, 3H, J=6.8 Hz), 1.23–
1.48 (m, 11H), 1.54–1.78 (m, 3H), 1.90–2.00 (m, 2H),
2.11–2.19 (m, 1H), 2.38* (dd, 1/5H, J=3.9, 16.6 Hz),
2.50 (t, 4/5H, J=12.7 Hz), 2.76 (d, 4/5H, J=12.7 Hz),
2.81* (s, 3/5H), 2.92* (d, 1/5H, J=6.8 Hz), 2.95 (s, 12/
5H), 3.00* (dd, 1/5H, J=4.9, 16.6 Hz), 3.74 (dd, 4/5H,
J=2.9, 18.6 Hz), 3.88–4.00 (m, 2H), 4.03–4.14 (m, 6/
5H), 4.24–4.35 (m, 9/5H), 4.32–4.35 (m, 1H), 4.42–4.47
(m, 1H), 4.45 (dd, 1H, J=5.4, 8.3 Hz), 4.71* (dd, 1/5H,
J=4.4, 9.4 Hz), 4.91 (dd, 4/5H, J=3.4, 9.4 Hz), 5.11 (d,
4/5H, J=7.8 Hz), 5.20–5.24 (m, 1H), 5.30–5.34* (m, 1/
5H), 6.64* (d, 1/5H, J=8.8 Hz), 6.99 (d, 4/5H, J=7.8
Hz), 7.12 (d, 4/5H, J=9.8 Hz), 7.24 (s, 4/5H), 7.45–
7.47* (m, 1/5H), 7.54* (t, 1/5H, J=5.4 Hz), 7.57–7.61*
(br, 1/5H, J=3.9 Hz), 7.65 (d, 4/5H, J=7.8 Hz); 13C
NMR (125 MHz, CDCl3, both rotamers) d ppm: 11.6,
11.8, 13.98, 14.04, 14.8, 15.3, 18.4, 21.3, 21.8, 22.48,
22.53, 23.4, 24.5, 24.7, 24.9, 25.2, 27.4, 28.9, 29.0, 29.5,
31.5, 31.6, 32.5, 35.1, 36.4, 37.6, 40.1, 40.5, 56.0, 58.0,
59.0, 60.9, 66.9, 71.5, 170.4, 170.9, 171.2, 171.5, 171.7,
172.1, 174.5; IR (KBr) cmÀ1: 3371, 2959, 2931, 2872,
1742, 1657, 1518, 1466, 1385, 1204; HRMS calcd for
C31H56N5O9 (M+H)+ 642.4078, found 642.4086. Anal.
.
Anal. calcd for C33H59N5O9 3/5H2O: C, 58.23; H, 8.91;
N, 10.29. Found: C, 57.93; H, 8.57; N, 9.99; [a]2D6 +56.0
(c 2.08, CHCl3).
4.2.9. Proline-substituted globomycin (1l). 98%, a color-
ꢀ
less solid (mp134–136 C), 1H NMR [500 MHz, CDCl3,
two rotamers (major/minor=5.2/1)] d ppm: 0.83 (d, 3H,
J=7.0 Hz), 0.87 (t, 3H, J=7.1 Hz), 0.95 (t, 3H, J=7.4
Hz), 1.09* (d, 1H, J=6.9 Hz), 1.13* (d, 1H, J=6.1 Hz),
1.22–1.38 (m, 15H), 1.49–1.59 (m, 1H), 1.62–1.66 (m,
1H), 1.89–1.93* (m, 1/6H), 1.95–2.02 (m, 5/6H), 2.02–
2.22 (m, 17/6H), 2.23–2.32* (m, 1/6H), 2.43–2.49 (m, 5/
6H), 2.51–2.53* (m, 1/6H), 2.86–2.95 (m, 2H), 3.49–3.54
(m, 5/6H), 3.57–3.62 (m, 1H), 3.64–3.79 (m, 13/6H),
3.92 (dd, 5/6H, J=5.9, 14.8 Hz), 3.96 (dd, 5/6H, J=5.4,
14.8 Hz), 4.07–4.13* (m, 1/6H), 4.21-4.32 (m, 14/6H),
4.37 (t, 5/6H, J=7.8 Hz), 4.54–4.56 (m, 10/6H), 4.74 (d,
5/6H, J=8.2 Hz), 4.93* (dd, 1/6H, J=4.8, 8.2 Hz),
5.05–5.13 (m, 1H), 5.20–5.21* (m, 1/6H), 5.41* (br d, 1/
6H, J=10.2 Hz), 5.55* (d, 1/6H, J=9.2 Hz), 5.76 (d, 5/
6H, J=5.8 Hz), 7.07* (d, 1/6H, J=3.8 Hz), 7.52 (br s,
5/6H), 7.56 (d, 5/6H, J=7.0 Hz), 7.78 (d, 5/6H, J=6.8
Hz), 8.34* (br, 1/6H), 8.75* (br d, 1/6H, J=8.2 Hz); 13C
NMR (125 MHz, CDCl3, 33 mM, both rotamers) d
ppm: 11.6, 11.9, 13.6, 13.9, 14.0, 14.4, 15.2, 18.0, 19.7,
22.4, 22.5, 24.7, 24.9, 25.1, 25.4, 26.4, 27.0, 28.87, 28.90,
29.0, 29.6, 31.5, 31.6, 31.9, 32.5, 34.6, 35.1, 40.9, 41.4,
42.3, 42.8, 48.0, 48.7, 53.9, 55.8, 56.6, 58.3, 58.9, 59.3,
61.7, 62.2, 62.8, 66.2, 66.4, 75.6, 76.4, 169.3, 169.5,
170.3, 170.7, 171.3, 171.4, 172.3, 172.8, 173.0, 173.8,
175.1; IR (KBr) cmÀ1: 3339, 2961, 2931, 2875, 1739,
1658, 1528, 1463, 1266, 1077; HRMS calcd for
C30H51N5O9Na (M+Na)+ calcd 648.3584, found
.
calcd for C31H55N5O9 27/50H2O: C, 57.15; H, 8.68; N,
10.75; Found: C 57.54; H, 8.69; N, 10.35; [a]2D5 À61.8 (c
1.13, CHCl3).
4.2.12. 5-Aza-globomycin (1o). 94%, a colorless solid
(mp212–213.5 ꢀC), 1H NMR (500 MHz, CD3OD,
major conformer) d ppm: 0.86–1.04 (m, 17H), 1.07 (d,
3H, J=6.8 Hz), 1.24 (d, 3H, J=6.8 Hz), 1.11–1.46 (m,
9H), 1.56–1.60 (m, 1H), 1.61–1.70 (m, 1H), 1.73–1.78
(m, 1H), 1.93–1.99 (m, 1H), 2.25–2.30 (m, 1H), 3.14–
3.17 (m, 1H), 3.25 (s, 3H), 3.82 and 3.88 (AB type d’s,
each 1H, J=17.1 Hz), 3.86 (dd, 1H, J=3.9, 11.0 Hz),
3.95–4.02 (m, 1H), 4.07 (dd, 1H, J=3.9, 11.0 Hz), 4.09–
4.12 (m, 1H), 4.13–4.18 (m, 1H), 4.22 (d, 1H, J=5.9
Hz), 4.35 (br d, 1H, J=5.9 Hz), 4.60 (t, 1H, J=3.9 Hz),
.
648.3589. Anal. calcd for C30H51N5O9 H2O: C, 55.97;
H, 8.30; N, 10.88, found: C, 56.12; H, 8.11; N, 10.87;
[a]2D5 À40.4 (c 1.14, CHCl3).

T. Kiho et al. / Bioorg. Med. Chem. 12 (2004) 337–361
361
13C NMR (125 MHz, CD3OD, both rotamers) d ppm:
References and notes
8.8, 11.5, 12.4, 12.5, 16.5, 19.0, 20.7, 20.8, 23.5, 24.4,
24.8, 27.0, 29.9, 33.3, 35.0, 37.5, 40.6, 45.5, 45.9, 46.1,
46.3, 50.4, 53.2, 58.3, 61.0, 64.7, 168.9, 169.7, 169.9,
170.38, 180.39, 170.7, 175.7; IR (KBr) cmÀ1: 3342, 2959,
2930, 2873, 1657, 1520, 1467, 1408, 1271, 1076; HRMS
calcd for C32H58N6O8Na (M+Na)+ 677.4214, foundd
677.4211. Anal. calcd for C32H58N6O8 1/3H2O: C,
58.16; H, 8.95; N, 12.72; Found: C 58.00; H, 8.81; N,
12.57; [a]2D4 À65.5 (c 1.05, CH3OH).
1. (a) Taxonomy of producing organism and fermentation.
Inukai, M.; Enokita, R.; Torikata, A.; Nakahara, M.;
Iwado, S.; Arai, M. J. Antibiot. 1978, 31, 410. (b) Isola-
tion and physico-chemical and biological characteriza-
tion: Inukai, M.; Nakajima, M.; Osawa, M.; Haneishi, T.;
Arai, M. J. Antibiot. 1978, 31, 421. (c) Structual determi-
nation of 1a: Nakajima, M.; Inukai, M.; Haneishi, T.;
Terahara, A.; Arai, M. J. Antibiot. 1978, 31, 426. (d)
Kogen, H.; Kiho, T.; Nakayama, M.; Furukawa, Y.;
Kinoshita, T.; Inukai, M. J. Am. Chem. Soc. 2000, 122,
10214. (e) Kiho, T.; Nakayama, M.; Kogen, H. Tetra-
hedron 2003, 59, 1685. (f) Kiho, T.; Nakayama, M.;
Yasuda, K.; Miyakoshi, S.; Inukai, M.; Kogen, H. Bioorg.
Med. Chem. Lett. 2003, 13, 2315.
.
4.2.13. 4-epi-Globomycin (1p). 95%, a colorless solid,
ꢀ
1
(mp, 81 C), H NMR [500 MHz, CDCl3, 16 mM, two
rotamers (major/minor=2.6/1)] d ppm: 0.77 (d, 6/7H,
J=6.8 Hz), 0.83–1.01 (m, 99/7H), 1.13–1.40 (m, 114/
7H), 1.46-1.53 (m, 5/7H), 1.55-1.64 (m, 9/7H), 1.68–1.74
(m, 5/7H), 1.77–1.88 (m, 1H), 1.94–1.99 (m, 5/7H),
2.13–2.17* (m, 2/7H), 2.24–2.27 (m, 1H), 2.84 (s, 6/7H),
2.94–2.98 (m, 5/7H), 3.12–3.18* (m, 2/7H), 3.18 (s, 15/
7H), 3.39 (br s, 5/7H), 3.51–3.56* (m, 2/7H), 3.61 (t, 5/
7H, J=7.8 Hz), 3.78–3.83 (m, 1H), 3.89–4.03 (m, 19/
7H), 4.10–4.19 (m, 2H), 4.27–4.31 (m, 1H), 4.41–4.44
(m, 9/7H), 4.54* (dd, 2/7H, J=3.9, 9.8 Hz), 4.60* (dd,
2/7H, J=3.9, 9.8 Hz), 4.77 (br s, 5/7H), 5.22* (t, 2/7H,
J=9.3 Hz), 5.27–5.29 (m, 5/7H), 6.36* (d, 2/7H, J=8.8
Hz), 7.23–7.25* (m, 2/7H), 7.42–7.45 (m, 1H), 7.50 (br s,
5/7H), 7.82–7.84 (m, 12/7H); 13C NMR (125 MHz,
CDCl3, both rotamers) d ppm: 11.3, 11.5, 11.6, 13.9,
14.0, 14.4, 15.3, 19.4, 19.7, 22.2, 22.4, 22.47, 22.50, 22.6,
23.0, 25.0, 25.3, 25.7, 25.9, 26.8, 26.9, 28.8, 29.0, 29.6,
29.7, 31.5, 31.6, 33.1, 35.4, 37.1, 37.7, 38.4, 39.6, 39.8,
40.9, 41.2, 41.4, 54.3, 56.5, 56.8, 57.0, 59.0, 59.6, 59.9,
61.4, 66.7, 67.0, 68.4, 73.9, 77.9, 168.6, 169.5, 170.5,
170.9, 171.0, 171.2, 171.3, 173.1, 174.4, 175.2, 176.1; IR
(CHCl3) cmÀ1: 3390, 3336, 2963, 2932, 2874, 1737, 1674,
1531, 1501, 1263; HRMS calcd for C32H57N5O9Na
(M+Na)+ 678.4054, found 678.4055. Anal. calcd for
2. (a) Omoto, S.; Suzuki, H.; Inouye, S. J. Antibiot. 1979, 32,
83. (b) Omoto, S.; Ogino, H.; Inouye, S. J. Antibiot. 1981,
34, 1416.
3. Pugsley, A. P. Microbiol. Rev. 1993, 57, 50.
4. (a) Inukai, M.; Takeuchi, M.; Shimizu, K.; Arai, M. J.
Antibiot. 1978, 31, 1203. (b) Hussain, M.; Ichihara, S.;
Mizushima, S. J. Biol. Chem. 1980, 255, 3707. (c) Dev,
I. K.; Harvey, R. J.; Ray, P. H. J. Biol. Chem. 1985, 260,
5891. (d) Ichihara, S.; Hussain, M.; Mizushima, S. J. Biol.
Chem. 1981, 256, 3125. (e) Witke, C.; Gotz, F. FEMS
Microl. Lett. 1995, 126, 233. (f) Braun, B., Wu, H. C. In
Bacterial Cell Wall; Ghuysen, J. M., Hakenbeck, R., Eds.;
Elsevier: Amsterdam, 1994; Chapter 14.
5. (a) Cavard, D. Arch. Microbiol. 1998, 171, 50. (b) Ochs-
ner, U. A.; Vasil, A. I.; Johnson, Z.; Vasil, M. L. J. Bac-
teriol. 1999, 181, 1099. (c) Paitan, Y.; Orr, E.; Ron, E. Z.;
Rosenberg, E. J. Bacteriol. 1999, 181, 5644. (d) Tibor, A.;
Decelle, B.; Letteson, J.-J. Infect. Immun. 1999, 67, 4960.
6. Abiko, A.; Liu, J.-F.; Masamune, S. J. Am. Chem. Soc.
1997, 119, 2586.
7. (a) Evans, D. A.; Bartroli, J.; Shih, T. L. J. Am. Chem.
Soc. 1981, 103, 2127. (b) Evans, D. A.; Dow, R. L.
Tetrahedron Lett. 1986, 27, 1007. (c) Rychnovsky, S. D.;
Buckmelter, A. J.; Dahanukar, V. H.; Skalitzky, D. J.
J. Org. Chem. 1999, 64, 6849.
8. Samuelsen, B.; Odham, G. Acta Chem. Scand. 1970, 24, 468.
9. Kitamura, M.; Tokunaga, M.; Ohkuma, T.; Noyori, R.
Org. Syn. 1992, 71, 1.
.
C32H57N5O9 H2O: C, 57.04; H, 8.83; N, 10.39, found:
C 56.77; H, 8.91; N, 10.24; [a]2D4 À1.23 (c 1.02,
CHCl3).
10. Coste, J.; Le-Nguyen, D.; Castro, B. Tetrahedron Lett.
1990, 31, 205.
11. Shioiri, T.; Yokoyama, Y.; Kasai, Y.; Yamada, S. Tetra-
hedron 1976, 32, 2211.
4.2.14. 8-N-Me-globomycin (1q). 94%, a colorless solid
(94–96 ꢀC), H NMR (500 MHz, CDCl3, 16 mM, major
1
conformer) d ppm: 0.86–0.99 (m, 15H), 1.16–1.20 (m,
6H), 1.22–1.38 (m, 10H), 1.47–1.60 (m, 2H), 1.67–1.74
(m, 1H), 1.81–1.87 (m, 1H), 2.12–2.18 (m, 1H), 2.25-
2.31 (m, 1H), 3.00 (s, 3H), 3.16–3.19 (m, 2H), 3.24 (br s,
1H), 3.28 (s, 3H), 3.57–3.60 (m, 1H), 3.70 (d, 1H,
J=18.5 Hz), 3.72-3.77 (m, 1H), 3.93-3.98 (m, 1H), 4.14–
4.18 (d, 2H, J=4.9 Hz), 4.36–4.41 (m, 2H), 4.55 (d, 1H,
J=18.5 Hz), 5.11–5.15 (m, 1H), 7.21 (d, 1H, J=6.8 Hz),
7.47–7.53 (br m, 2H); 13C NMR (125 MHz, CDCl3, 30
mM, major conformer) d ppm: 11.8, 12.6, 14.0, 14.1,
19.5, 22.0, 22.6, 23.0, 25.2, 25.4, 27.0, 29.1, 31.0, 31.7,
35.5, 35.9, 38.1, 40.6, 41.4, 51.9, 54.5, 54.8, 56.8, 62.0,
67.8, 68.0, 77.3, 168.7, 170.2, 171.4, 172.2, 172.6, 177.1;
IR (KBr) cmÀ1: 3370, 2958, 2930, 2873, 1739, 1654, 1530,
1504, 1468, 1206; HRMS calcd for C33H59N5O9Na
(M+Na)+ calcd 692.4211, found 692.4185. Anal. calcd
12. Boden, E. P.; Keck, G. E. J. Org. Chem. 1985, 50, 2394.
13. (a) Carpino, L. A.; El-Faham, A. J. Org. Chem. 1994, 59,
695. (b) Carpino, L. A. J. Am. Chem. Soc. 1993, 115,
4397. (c) Carpino, L. A.; El-Faham, A. J. Org. Chem.
1995, 60, 3561. (d) Humphrey, J. M.; Chamberlin, A. R.
Chem. Rev. 1997, 97, 2243.
14. The cyclization yield for 1m was very low.
15. Compound 1j and 1o are insoluble in CDCl3.
16. O-Bn globomycin derivative in l-Ser residue was also
inactive against Escherichia coli SANK 70569 (>50 mg/
mL). Diacetylation of 1a in l-Ser and l-allo-Thr residue
diminished the activity.2b
17. Kiho, T.; Iwata, Y.; Kogen, H.; Miyamoto, S. Con-
formational analysis of globomycin with a signal pepti-
dase II inhibitory activity using molecular dynamics
simulation. DrugDes. Discov. , in press.
18. National Committee for Clinical Laboratory Standards.
Standard Methods for Dilution Antimicrobial Susceptibility
Tests for Bacteria that Grow Aerobically; Approved Stan-
dard (M7-A5); NCCLS: Villanova, 2000; p7.
.
for C33H59N5O9 H2O: C, 57.62; H, 8.94; N, 10.18,
found: C, 57.71; H, 8.94; N, 10.04; [a]2D6 À44.5 (c 0.50,
CH3OH).