Discovery of Leucyladenylate Sulfamates as Novel Leucyl-tRNA Synthetase (LRS)-Targeted Mammalian Target of Rapamycin Complex 1 (mTORC1) Inhibitors

Article abstract of DOI:10.1021/acs.jmedchem.6b01190

Recent studies indicate that LRS may act as a leucine sensor for the mTORC1 pathway, potentially providing an alternative strategy to overcome rapamycin resistance in cancer treatments. In this study, we developed leucyladenylate sulfamate derivatives as LRS-targeted mTORC1 inhibitors. Compound 18 selectively inhibited LRS-mediated mTORC1 activation and exerted specific cytotoxicity against colon cancer cells with a hyperactive mTORC1, suggesting that 18 may offer a novel treatment option for human colorectal cancer.

Full text of DOI:10.1021/acs.jmedchem.6b01190

Subscriber access provided by TUFTS UNIV
Brief Article
Discovery of Leucyladenylate Sulfamates as Novel
Leucyl-tRNA Synthetase (LRS)-targeted Mammalian
Target of Rapamycin Complex 1 (mTORC1) Inhibitors
Suyoung Yoon, Jong Hyun Kim, Sung-Eun Kim, Changhoon Kim, Jihyae Ann, Yura Koh, Jayun Jang,
Sungmin Kim, Hee-sun Moon, Won Kyung Kim, Sang Kook Lee, Jiyoun Lee, Sunghoon Kim, and Jeewoo Lee
J. Med. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jmedchem.6b01190 • Publication Date (Web): 19 Oct 2016
Downloaded from http://pubs.acs.org on October 20, 2016
Just Accepted
“Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted
online prior to technical editing, formatting for publication and author proofing. The American Chemical
Society provides “Just Accepted” as a free service to the research community to expedite the
dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts
appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been
fully peer reviewed, but should not be considered the official version of record. They are accessible to all
readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered
to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published
in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just
Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor
changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers
and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors
or consequences arising from the use of information contained in these “Just Accepted” manuscripts.
Journal of Medicinal Chemistry is published by the American Chemical Society. 1155
Sixteenth Street N.W., Washington, DC 20036
Published by American Chemical Society. Copyright © American Chemical Society.
However, no copyright claim is made to original U.S. Government works, or works
produced by employees of any Commonwealth realm Crown government in the course
of their duties.

Page 1 of 7
Journal of Medicinal Chemistry
1
2
3
4
5
6
7
8
9
Discovery of Leucyladenylate Sulfamates as Novel Leucyl-
tRNA Synthetase (LRS)-targeted Mammalian Target of Ra-
pamycin Complex 1 (mTORC1) Inhibitors
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Suyoung Yoon,† Jong Hyun Kim,‡ SungꢀEun Kim,† Changhoon Kim,† Jihyae Ann,† Yura Koh,† Jayun
Jang,‡ Sungmin Kim,‡ Heeꢀsun Moon,‡ Won Kyung Kim,† Sangkook Lee,† Jiyoun Lee,§ Sunghoon
Kim, ‡,∥ Jeewoo Lee*,†
† Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 151ꢀ742, Korea
‡ Medicinal Bioconvergence Research Center, College of Pharmacy, Seoul National University, Seoul 151ꢀ742, Korea
∥ Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Techꢀ
nology, Seoul National University, Seoul 151ꢀ742, Korea
§ Department of Global Medical Science, Sungshin University, Seoul 142ꢀ732, Korea
ABSTRACT: Recent studies indicate that LRS may act as a leucine sensor for the mTORC1 pathway, potentially providing an
alternative strategy to overcome rapamycinꢀresistance in cancer treatments. In this study, we developed leucyladenylate sulfamate
derivatives as LRSꢀtargeted mTORC1 inhibitors. Compound 18 selectively inhibited LRSꢀmediated mTORC1 activation and exertꢀ
ed specific cytotoxicity against colon cancer cells with a hyperactive mTORC1, suggesting that compound 18 may offer a novel
treatment option for human colorectal cancer.
mTORC1 activity, and have the potential to overcome raꢀ
pamycinꢀresistance in cancer treatment.
■
INTRODUCTION
Mammalian target of rapamycin (mTOR) is a multiꢀdomain
kinase that plays a central role in the regulation of cell growth,
proliferation, metabolism and autophagy. mTOR consists of
two structurally and functionally distinct protein complexes,
mTORC1 activity is regulated by various environmental sigꢀ
nals, such as the levels of nutrients, energy, and oxygen. Altꢀ
hough the complete mechanism of how mTORC1 senses these
signals still remains a mystery, the proteinogenic amino acid
leucine is considered to be the master controller in amino acidꢀ
dependent mTORC1 signaling.13, 14 More importantly, several
recent studies have reported that leucylꢀtRNA synthetase
(LRS) may act as an intracellular leucine sensor by directly
binding to RagD GTPase, one of the key mediators of the
amino acidꢀdependent mTORC1 pathway.15, 16 LRS is a memꢀ
ber of the class I aminoacylꢀtRNA synthetase (ARSs) family
that catalyzes the ATPꢀdependent ligation of amino acids to
cognate transfer RNA (tRNA) in protein biosynthesis. Because
of their pivotal role in cell survival, ARSs have been effective
targets for antibiotics to overcome bacterial resistance,17ꢀ20 and
also have been studied as antiꢀcancer agents.21ꢀ23 In addition to
its traditional role, LRS appears to participate in mTORC1
activation by acting as a GTPaseꢀactivating protein (GAP) for
Rag GTPase in a leucineꢀdependent manner. Moreover, the
leucineꢀinduced activation of mTORC1 can be inhibited by
leucine analogues, such as leucinol, without affecting the leuꢀ
cine charging ability of LRS, suggesting that LRSꢀtargeted
inhibitors can suppress mTORC1 activity.15, 24, 25
mTOR complex 1 (mTORC1) and mTOR complex
2
(mTORC2).1 In particular, mTORC1 regulates protein syntheꢀ
sis by phosphorylating the two major substrates, S6 kinase 1
(S6K1) and the translational regulators eukaryotic translation
initiation factor 4E (elF4E)ꢀbinding protein 1 (4EꢀBP1), both
of which are known to play crucial roles in cell proliferation
and tumorigenesis.2ꢀ4 Therefore, much research effort has foꢀ
cused on the development of compounds that suppress the
kinase activity of mTORC1 and act as antiꢀcancer agents.5ꢀ7
The most studied compounds are rapamycin and its derivatives
that directly bind near the mTOR kinase domain by forming a
complex with the FK506ꢀbinding protein 12 (FKBP12);
temsirolimus and everolimus have been approved by the FDA
9
for the treatment of advanced renal cell carcinoma,8, and
ridaforolimus showed a promising outcome in a recent clinical
trial for advanced soft tissue and bone sarcoma.10, 11 Unfortuꢀ
nately, the efficacy of these rapamycin analogs for the treatꢀ
ment of various cancer patients has been generally disappointꢀ
ing, mainly because these agents are cytostatic and only parꢀ
tially inhibit mTORC1 activity.7, 12 Although rapamycin is a
highly specific allosteric inhibitor of mTORC1, the mTORC1
pathway involves multiple regulatory mechanisms and comꢀ
plex feedback loops that have not yet been fully understood,
thus resulting in an incomplete inhibition of kinase activity.
Therefore, small molecules targeting other possible regulators
may offer an alternative strategy for the suppression of
Previously, we have demonstrated that a leucine analog, (S)ꢀ
4ꢀisobutyloxazolidinꢀ2ꢀone, selectively inhibited downstream
phosphorylation of mTORC1 by blocking the leucineꢀsensing
ability of LRS.26 Furthermore, this analog exhibited cytotoxiꢀ
city against rapamycinꢀresistant colon cancer cells without
affecting the catalytic activity of LRS, indicating that LRS has
the potential to serve as a novel therapeutic target. In a continꢀ
ACS Paragon Plus Environment

Journal of Medicinal Chemistry
uing effort to develop LRSꢀtargeted mTORC1 inhibitors, we
Page 2 of 7
figuration was confirmed by following previously described
procedures.33 Amide coupling between compound 10 and the
protected chiral acid produced compound 11, four acetyl
groups of which were then removed in the presence of sodium
methoxide to yield the final compound 12.
1
2
3
4
5
6
7
8
designed a new series of compounds based on the structure of
aminoacyl adenylate 1, an enzymatic reaction intermediate of
ARSs (Figure 1). Aminoacyl adenylates have been extensiveꢀ
ly studied as a lead compound for ARS inhibitors.27, 28 Specifiꢀ
cally, replacing the hydrolytically unstable acylphosphate
group with its isosteres, such as acylsulfamate, resulted in
chemically stable and potent inhibitors with IC50 values in the
submicromolar to low micromolar range versus the correꢀ
sponding ARSs.29ꢀ31 Based on these previous findings, we
synthesized a library of leucyladenylate sulfamates by modifyꢀ
ing adenine (group 1), ribose (group 2) and leucine moieties
(group 3) and evaluated their biological activity. When we
tested these compounds by using immunoblots, we found that
group 1 compounds with a lipophilic R2 group generally
showed good activity against mTORC1, while R2 = I derivaꢀ
tives were most active. On the other hand, any modifications
in the ribose moiety (group 2) lead to the loss of activity.
Among the group 3 compounds, replacement of the αꢀamino
group of leucine (R1) with a hydroxyl group significantly enꢀ
hanced inhibitory activity against mTORC1, while introducꢀ
tion of an additional hydroxyl group also resulted in good inꢀ
hibitory effect. Among the more than 70 compounds that we
tested, herein we report three representative compounds deꢀ
scribed in Figure 1. Each of these three compounds contains
distinct structural features that are required for either LRS
inhibition or LRSꢀmediated mTORC1 suppression. In this
paper, we describe their syntheses, and inhibitory effects on
the mTORC1 pathway together with their leucylation activity.
We also evaluated cancer cellꢀspecific cytotoxicity of these
analogs in various types of cancer cells.
Scheme 1. Synthesis of compound 6a
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
aReagents & conditions: (a) i) NaH, THF, ii) NH2SO2Cl, THF;
(b) NꢀBoc leucine, DCC, DMAP, CH2Cl2; (c) 80% aq. TFA.
Scheme 2. Synthesis of compound 12 a
aReagents & conditions: (a) MMTCl, pyridine, DMF; (b) Ac2O,
NEt3, DMAP, CH3CN; (c) 80% aq. AcOH; (d) i) NaH, THF, ii)
NH2SO2Cl, THF; (e) chiral acid, DCC, DMAP, CH2Cl2; (f) Naꢀ
OMe in MeOH.
Scheme 3. Synthesis of compound 18 a
Figure 1. Amionacyl adenylate 1 and leucyladenylate sulfamates
■ RESULTS AND DISCUSSION
Chemistry. Synthesis of leucyladenylate sulfamate 6 began
with commercially available 2’,3’ꢀisopropylidene adenosine 3
as shown in Scheme 1. Compound 3 was reacted with sulꢀ
famoyl chloride prepared in a quantitative yield from
chlorosulfonyl isocyanate according to previously described
procedures.32 Sulfamoylated adenosine 4 was then coupled
with NꢀBocꢀleucine to give the leucine adduct 5, which was
subsequently hydrolyzed to leucyladenylate sulfamate 6.
Synthesis of a synꢀdiol analog of leucyladenylate sulfamate
12 was carried out as illustrated in Scheme 2. 5’ꢀOꢀ
Monomethoxytritylation (MMT) of compound 7 followed by
acetylation of adenosine provided the fully protected adenoꢀ
sine 8. Selective removal of MMT group and subsequent sulꢀ
famoylation produced the sulfamate intermediate 10. (2S,3R)ꢀ
dihydroxyꢀ4ꢀmethylpentanoic acid (DMPA) protected by Oꢀ
diacetyl was prepared from 4ꢀmethylꢀ2ꢀpentenoic acid in 4
steps by Sharpless asymmetric dihydroxylation, and its conꢀ
aReagents & conditions: (a) Ac2O, NEt3, DMAP, CH3CN; (b)
POCl3, Et4NCl, PhNMe2, CH3CN; c) isoamyl nitrite, I2, CuI,
CH2I2, THF; (d) 7M NH3 in MeOH; (eꢀj) the same conditions as
(aꢀf) in scheme 2
Synthesis of a hydroxyl surrogate of leucyladenylate sulfaꢀ
mate 18 is shown in Scheme 3. 2ꢀIodoadenosine (14) was
synthesized from guanosine (13) in 4 steps according to previꢀ
ACS Paragon Plus Environment

Page 3 of 7
Journal of Medicinal Chemistry
ously described procedures.34 Compound 14 was converted to
mediated mTORC1 activation is independent of the leucylaꢀ
tion activity of LRS on its cognate tRNA, which is again in
1
2
3
4
5
6
7
the corresponding 5’ꢀOꢀsulfamoylated intermediate 16 by folꢀ
lowing the route described in Scheme 2. (2S)ꢀ
hydroxyisocaproic acid (HICA, Lꢀleucic acid) protected by Oꢀ
acetyl was prepared by acetylation of commercially available
Lꢀleucic acid. A peptide coupling reaction between compound
16 and OꢀacetylꢀLꢀleucic acid followed by deprotection of the
acetyl group provided the final compound 18.
26
agreement with previously reported observations.15,
More
interestingly, the side chain modifications of an αꢀhydroxyl
group in the leucyl side chain and a 2ꢀiodo group in the adeꢀ
nine, both of which adversely affected the catalytic activity of
LRS, resulted in favorable effects for the mTORC1 inhibition
Biological Activity. Because LRS catalyzes leucylation reꢀ
action with its cognate tRNA during protein synthesis, we first
assessed the inhibitory effects of compounds 6, 12 and 18 on
the catalytic activity of LRS by performing aminoleucylation
assays to determine the IC50 values. Given that the previously
reported acylsulfamate adenylates were found to be potent
inhibitors of the corresponding ARSs, these compounds would
be likely to exhibit inhibitory effects of LRS. As shown in
Figure 2, compound 6 proved to be a potent inhibitor of LRS
with an IC50 value of 22.34 nM. However, compounds 12 and
18 inhibited LRS to a much lesser extent, showing 3ꢀ and 15ꢀ
fold higher IC50 values (70.04 nM and 337.1 nM, respectively)
than compound 6. Replacement of the αꢀamino group of leuꢀ
cine side chain with a hydroxyl group (12), and the introducꢀ
tion of a 2ꢀiodo group to adenine (18) weakened the overall
binding affinity toward the LRS active site. Moreover, considꢀ
ering foldꢀchanges between compounds, we conclude that
modification of the adenine moiety has a higher impact on the
catalytic activity than the modification of the leucyl side chain.
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
Figure 3. Doseꢀdependent inhibition of leucineꢀinduced mTORC1
activation of compounds 6, 12 and 18 in HEK293 cells
To further investigate the mechanisms of these sulfamates in
LRSꢀmediated mTORC1 activation, we performed in vitro
mTORC1 kinase assays using purified mTOR. As demonstratꢀ
ed in Figure 4, compounds 12 and 18 both did not inhibit the
kinase activity of the purified mTOR at 200 µM, suggesting
that these compounds are highly selective toward LRS, and
did not act as an ATPꢀcompetitive inhibitor of the mTOR
complex. We believe that these two sulfamates, 12 and 18,
inhibited the mTORC1 activity by selectively blocking the
LRSꢀmediated mTORC1 activation pathway, rather than diꢀ
rectly interacting with mTOR.
Figure 2. Inhibitory activities of compounds 6, 12 and 18 for
catalytic leucylation.
Next, we determined the effects of compounds 6, 12, and 18
on leucineꢀinduced mTORC1 activation using the immunobꢀ
lotting method. In our previous study, pretreatment of leucinol
analogs blocked leucineꢀinduced phosphorylation of S6K, an
mTORC1 substrate, by directly interacting with LRS26; beꢀ
cause the newly designed compounds have additional adenoꢀ
sine moieties, we would expect to observe stronger inhibitory
effects for S6K phosphorylation. We pretreated HEK293 cells
with each compound at different concentrations as well as
with rapamycin as a control at 100 nM, and then activated
mTORC1 by treating the cells with leucine for 10 min. As
shown in Figure 3, pretreatment of rapamycin blocked phosꢀ
phorylation of S6K, whereas pretreatment of compound 6 did
not affect S6K phosphorylation at all (Figure 3A). Notably,
compounds 12 and 18 showed a doseꢀdependent inhibition of
mTORC1 in HEK293 cells (Figure 3B, 3C) while compound
18 appeared to be more potent than compound 12. Interestingꢀ
ly, these three sulfamates inhibited leucineꢀinduced mTORC1
activation in the order of 18 > 12 > 6, in contrast to the order
of their inhibitory effects for the catalytic reaction (6 > 12 >
18). This apparently opposite trend suggests that LRSꢀ
Figure 4. Effects of compounds 12 and 18 on the kinase activity
of the purified mTOR.
Finally, to examine the antiꢀcancer activity of leucyladenylꢀ
ate sulfamates, we performed the sulforhodamine B (SRB)
colorimetric assays for cytotoxicity.35 We treated compounds
6, 12, and 18 with six different types of cancer cell lines toꢀ
gether with etoposide as a positive control. To determine canꢀ
cer cell selectivity, we also treated all four compounds with
normal human lung epithelial cells (MRCꢀ5), with the measꢀ
ured IC50 values shown in Table 1. Compound 6, the most
ACS Paragon Plus Environment

Journal of Medicinal Chemistry
Page 4 of 7
Table 1. Relative cell growth inhibition of compounds 6, 12 and 18 for various cancer cell types and normal cells. a
1
2
3
IC (ꢁM)
A549
HCT116
K562
MDAꢀMBꢀ231 SKꢀHEPꢀ1
SNU638
MRC5
50
0.59
1.64
1.75
0.24
0.24
1.44
0.54
1.25
0.4
0.73
7.01
12.6
7.35
0.54
6.81
5.63
0.25
0.84
8.98
5.7
5.9
>50
>50
12.7
6
12
4
5
6
7
2.22
1.06
1.79
18
8
Etoposide
0.56
9
aA549: lung cancer cells, MDAꢀMBꢀ231: breast cancer cells, SKꢀHepꢀ1: liver cancer cells, SNU638: stomach cancer cells, HCT116: coꢀ
lon cancer cells, K562: leukemia cells, MRC5: lung normal epithelial cell
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
to >95% purity, as determined by highꢀperformance liquid chromaꢀ
tography (HPLC). HPLC was performed on an Agilent 1120 Compact
LC (G4288A) instrument using an Agilent Eclipse Plus C18 column
(4.6 x 250 mm, 5 µm) and a Daicel Chiralcel ODꢀH column (4.6 x
250 mm, 5 µm).
potent LRS inhibitor, exhibited the greatest cytotoxicity that
was even greater than that of etoposide in all types of cells,
except A549 and SNU638. Given that LRS plays a crucial role
in protein synthesis, it is not surprising that compound 6 is
highly cytotoxic in both cancer cells and normal cells. In conꢀ
trast, compounds 12 and 18 showed selective cytotoxicity
against cancer cells. Specifically, compound 18 showed potent
cytotoxicity against colon cancer cells (HCT116) and leukeꢀ
mia cells (K562) while exhibiting much less cytotoxicity
against normal cells compared with compound 6 and etopoꢀ
side. This result is particularly promising because several reꢀ
cent studies have reported that hyperactive mTORC1 is one of
the distinctive features in human colorectal cancer.36, 37 sugꢀ
gesting that compound 18 exerted colon cancer specific cytoꢀ
toxicity by selective inhibition of mTORC1.
((3aR,4R,6R,6aR)ꢀ6ꢀ(6ꢀaminoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ2,2ꢀdimethyltetraꢀ
hydrofuro[3,4ꢀd][1,3]dioxolꢀ4ꢀyl)methyl sulfamate (4). Compound 4
was prepared by following the reported procedure.32 Yield 69%, white
1
solid; HꢀNMR (300MHz, CD3OD) δ 8.27 (s, 1H, CH), 8.22 (s, 1H,
CH), 6.24 (d, 1H, CH, J = 2.55 Hz), 5.42 (dd, 1H, CH, J = 6.24 Hz,
2.58 Hz), 5.13 (dd, 1H, CH, J = 6.21 Hz, 2.73 Hz), 4.51 (m, 1H, CH),
4.32 (dd, 1H, CH, J = 10.62 Hz, 4.59 Hz), 4.24 (dd, 1H, CH, J =
10.62 Hz, 5.31 Hz), 1.60 (s, 3H, CH3), 1.39 (s, 3H, CH3)
((3aR,4R,6R,6aR)ꢀ6ꢀ(6ꢀaminoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ2,2ꢀdimethyltetra
hydrofuro[3,4ꢀd][1,3]dioxolꢀ4ꢀyl)methyl ((tertꢀbutoxy carbonyl)ꢀLꢀ
leucyl)sulfamate (5). To a solution of compound 4 (0.54 mmol) in
anhydrous MC (25 mL) was added NꢀBoc leucine (0.81 mmol) and
DMAP (0.01 mmol) at 0oC. 1M DCC in MC (0.81 mmol) was added
dropwise, stirred for 2 h at room temperature. The solution was filꢀ
tered on celite pad, washed with EtOAc (20 mL), and then concenꢀ
trated. The filtrate was purified by column chromatography over silica
gel (EtOAc:MeOH=10:1) to give the compound 5 (0.46 mmol). Yield
■ CONCLUSION
In this study, we developed leucyladenylate sulfamate derivꢀ
atives that directly interact with LRS to inhibit the mTORC1
pathway. Compound 6 inhibited the catalytic activity of LRS
but did not affect the leucineꢀinduced mTORC1 activation,
whereas compound 18 inhibited mTORC1 activation, while it
also inhibited the catalytic activity of LRS to a much lesser
degree compared to compounds 6 and 12. Furthermore, both
compounds 12 and 18 did not affect the kinase activity of the
purified mTOR, indicating that the mTORC1ꢀspecific activity
of these compounds arose from blocking the leucineꢀsensing
ability of LRS rather than from interacting with mTOR directꢀ
ly. Cytotoxicity screening in various types of cancer cells and
normal cells revealed that compound 6 showed the greatest
cytotoxicity, probably due to a nonꢀspecific inhibition of LRS.
Compounds 12 and 18 demonstrated cytotoxicity against all
types of cancer cells but not against normal cells. Most notaꢀ
bly, compound 18 exerted highly specific cytotoxicity against
colon cancer cells that are known to have hyperactive
mTORC1. We believe that compound 18 may serve as a useꢀ
ful tool to study the role of LRS in the mTORC1 pathway but
may also offer a novel treatment option for human colorectal
cancer.
1
85%, white solid; HꢀNMR (300MHz, CD3OD) δ 8.47 (s, 1H, CH),
8.21 (s, 1H, CH), 6.22 (d, 1H, CH, J = 3.30 Hz), 5.33 (dd, 1H, CH, J
= 5.88 Hz, 3.48 Hz), 5.10 (d, 1H, CH, J = 5.67 Hz), 4.53 (m, 1H,
CH), 4.23 (d, 2H, CH2, J = 3.48 Hz), 4.06 (m, 1H, CH), 1.69 (m, 1H,
CH), 1.60 (s, 3H, CH3), 1.52 (m, 2H, CH2), 1.43 (s, 9H, CH3), 1.40 (s,
3H, CH3), 0.93 (dd, 6H, CH3, J = 6.60 Hz, 2.76 Hz)
((2R,3S,4R,5R)ꢀ5ꢀ(6ꢀaminoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ3,4ꢀdihydroxytetra
hydrofuranꢀ2ꢀyl)methyl (Lꢀleucyl) sulfamate trifluoroacetate salt (6).
The compound 5 (0.38 mmol) was dissolved in 80% aquous TFA
(2mL) and stirred for 2 h at room temperature. The reaction mixture
was evaporated and washed with EtOAc (20 mL). Aqueous layer was
concentrated under reduced pressure to give crude pale yellow solid.
The solid was purified by ionꢀexchange resin (HP20SS) to give the
compound 6 (0.16 mmol). Yield 42%, white solid; 1HꢀNMR
(500MHz, CD3OD) δ 8.49 (s, 1H, CH), 8.20 (s, 1H, CH), 6.07 (d, 1H,
CH, J = 5.30 Hz), 4.62 (t, 1H, CH, J = 5.10 Hz), 4.39ꢀ4.29 (m, 4H, , 2
CH, CH2), 3.61 (dd, 1H, CH, J = 8.50 Hz, 4.85 Hz), 1.78 (m, 2H,
CH2), 1.57 (m, 1H, CH), 0.95 (dd, 6H, CH3, J = 15.95 Hz, 6.00 Hz);
HRMSꢀFAB m/z [M+H]+ C16H25N7O7SH+ calcd 460.1536, Found:
460.4898.
(2R,3R,4R,5R)ꢀ2ꢀ(6ꢀaminoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ5ꢀ(((4ꢀmethoxyphenyl)
diphenylmethoxy)methyl)tetrahydrofuranꢀ3,4ꢀdiyl diacetate (8). 5’ꢀOꢀ
Monomethoxytrityl (MMT) protected adenosine (2.27 mmol) was
prepared by following the reported procedure.38 To a solution of 5’ꢀOꢀ
MMT protected adenosine (2.05 mmol) in acetonitrile (50 mL) at 0oC
was added DMAP (0.21 mmol), TEA (6.15 mmol) and acetic
anhydride (6.15 mmol). The mixture was stirred for 4hr at room temꢀ
perature. Aqueous portion was extracted with EtOAc (50 mL). The
organic phase was dried over MgSO4 and concentrated, which was
purified by column chromatography (EtOAc:MeOH = 40:1, v/v) to
give compound 8 (1.6 mmol). Yield 78%, white solid; 1HꢀNMR
(300MHz, CDCl3) δ 8.01 (S, 1H, CH), 7.90 (S, 1H, CH), 7.42 (m, 4H,
Ar), 7.32 (m, 4H, Ar), 7.25 (m, 4H, Ar), 6.18 (m, 4H, Ar), 6.26 (d,
1H, CH, J = 6.6 Hz), 6.07 (t, 1H, CH, J = 5.3 Hz), 5.67 (dd, 1H, CH,
■ EXPERIMENTAL SECTION
General Methods. All chemical reagents were commercially
available. Melting points were determined on a Büchi Melting Point
Bꢀ540 apparatus and are uncorrected. Silica gel column chromatogꢀ
raphy was performed on silica gel 60, 230ꢀ400 mesh, Merck. Nuclear
magnetic resonance (1HꢀNMR and 13CꢀNMR) spectra were recorded
on JEOL JNMꢀLA 300 [300 MHz (1H), 75 MHz (13C)] and Bruker
Avance 400 MHz FTꢀNMR [400 MHz (1H), 100 MHz(13C)] specꢀ
trometers. Chemical shifts are reported in ppm units with Me4Si as a
reference standard. Mass spectra were recorded on a VG Trioꢀ2 GCꢀ
MS and 6460 Triple Quad LC/MS. All final compounds were purified
ACS Paragon Plus Environment

Page 5 of 7
Journal of Medicinal Chemistry
1
J = 5.1 Hz, 3.1 Hz), 4.31 (m, 1H, CH), 3.78 (S, 3H, CH3), 3.46 (m,
white solid; HꢀNMR (300MHz, CDCl3) δ 8.31 (s, 1H, CH), 6.28 (d,
1H, CH, J = 6.69 Hz), 5.75 (m, 1H, CH), 5.54 (m, 1H, CH), 4.70 (m,
1H, CH), 4.42ꢀ4.34 (m, 2H, CH2), 2.14 (s, 3H, CH3), 1.98 (s, 6H,
CH3), 1.64 (m, 2H, CH2), 1.50 (m, 1H. CH), 0.79(dd, 6H, CH3, J =
13.17 Hz, 6.60 Hz)
1H, CH), 2.10 (S, 3H, CH3), 2.05 (s, 3H, CH3)
1
2
3
4
5
6
7
8
(2R,3R,4R,5R)ꢀ2ꢀ(6ꢀaminoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ5ꢀ(hydroxymethyl)
tetrahydrofuranꢀ3,4ꢀdiyl diacetate (9). 80% aquous AcOH (100 mL)
was slowly added to compound 8 (1.52 mmol) and the reaction
mixture was stirred for 12 h at room temperature. The mixture was
evaporated, neutralized with NaHCO3 and extrated with EtOAc (150
mL x 2). Organic layer was combined, dried over MgSO4 and evapoꢀ
rated. The residue was purified by column chromatography
(EtOAc:MeOH = 20:1, v/v) to give compound 9 (0.97 mmol). Yield
((2R,3S,4R,5R)ꢀ5ꢀ(6ꢀaminoꢀ2ꢀiodoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ3,4ꢀdihydroxy
tetrahydrofuranꢀ2ꢀyl)methyl
((S)ꢀ2ꢀhydroxyꢀ4ꢀmethylpentanoyl)
sulfamate (18). Compound 18 was prepared by following the
1
procedure described for compound 12. Yield 67%, white solid; Hꢀ
NMR (500MHz, CD3OD) δ 8.26 (s, 1H, CH), 5.98 (d, 1H, CH, J =
4.90 Hz), 4.55 (t, 1H, CH, J = 4.85 Hz), 4.50 (dd, 1H, CH, J = 11.20
Hz, 2.80 Hz), 4.44 (dd, 1H, CH, J = 11.2 Hz, 3.40 Hz), 4.36 (t, 1H,
CH, J = 4.70 Hz), 4.28 (q, 1H, CH, J = 3.60 Hz), 4.03 (t, 1H, CH, J =
6.30 Hz), 1.83ꢀ1.79 (m, 1H, CH), 1.52ꢀ1.47 (m, 2H, CH2), 0.90 (dd,
6H, CH3, J = 6.60 Hz, 2.25 Hz); HRMSꢀESI m/z [M+H]+
C16H23IN6O8SH+ calcd 587.0343, Found 587.0410.
1
64%, white solid; HꢀNMR (300MHz, CDCl3) δ 8.32 (s, 1H, CH),
9
7.85 (S, 1H, CH), 6.26 (S, 2H, NH2), 6.03 (d, 2H, CH2, J = 2.0 Hz),
5.70 (m, 1H, CH), 4.37 (s, 1H, CH), 3.99 (dd, 1H, CH, J = 13.0 Hz,
1.4 Hz), 3.86 (d, 1H, CH, J = 12.3 Hz), 2.18 (s, 3H, CH3), 2.02 (s,
3H, CH3)
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
(2R,3R,4R,5R)ꢀ2ꢀ(6ꢀaminoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ5ꢀ((sulfamoyloxy)
methyl)tetrahydrofuranꢀ3,4ꢀdiyl diacetate (10). Compound 10 was
prepared by following the procedure described for compound 4. Yield
■ AUTHOR INFORMATION
1
93%, white solid; HꢀNMR (300MHz, CDCl3) δ 8.34 (s, 1H, CH),
8.00 (s, 1H, CH), 6.16 (d, 1H, CH, J = 6.1 Hz), 5.91 (t, 1H, CH, J =
5.9 Hz), 5.72 (m, 1H, CH), 2.16 (s, 3H, CH3), 2.07 (s, 3H, CH3)
(2R,3R,4R,5R)ꢀ2ꢀ(6ꢀaminoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ5ꢀ(((Nꢀ((2S,3R)ꢀ2,3ꢀ
diacetoxyꢀ4ꢀmethylpentanoyl)sulfamoyl)oxy)methyl)tetrahydrofuranꢀ
3,4ꢀdiyl diacetate (11). Compound 11 was prepared by following the
Corresponding Author
* Phone, 82ꢀ2ꢀ880ꢀ7846; Eꢀmail, jeewoo@snu.ac.kr.
Notes
The authors declare no competing financial interest.
1
procedure described for compound 5. Yield 79%, white solid; Hꢀ
■ ACKNOWLEDGMENT
NMR (300MHz, CD3OD) δ 8.47 (s, 1H, CH), 8.34 (s, 1H, CH), 6.29
(d, 1H, CH, J = 5.3 Hz), 5.85 (t, 1H, CH, J = 5.49 Hz), 5.68 (dd, 1H,
CH, J = 5.67 Hz, 4.0 Hz), 5.14 (dd, 1H, CH, J = 8.8 Hz, 2.7 Hz), 5.07
(d, 1H, CH, J = 2.7 Hz), 4.52 (m, 2H, CH, NH), 3.68 (s, 1H, CH),
2.14 (s, 3H, CH3), 2.03 (s, 3H, CH3), 2.01 (s, 3H, CH3), 2.04 (m, 1H,
CH), 0.94 (d, 3H, CH3, J = 2.5 Hz), 0.91 (d, 3H, CH3, J = 2.4 Hz)
((2R,3S,4R,5R)ꢀ5ꢀ(6ꢀaminoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ3,4ꢀdihydroxytetra
This research was supported by the Global Frontier Project grant
(NRFꢀ2012M3A6A4054928) of National Research Foundation
funded by the Ministry of Education, Science and Technology of
Korea.
■ ABBREVIATIONS
hydrofuranꢀ2ꢀyl)methyl ((2S,3R)ꢀ2,3ꢀdihydroxyꢀ4ꢀmethylpentanoyl)
sulfamate (12). Compound 11 (0.056 mmol) was dissolved in 0.02 M
sodium methoxide solution in methanol (3 mL) and stirred for 2 h at
room temperature. DOWEX 50WX8 hydrogen form resin (10 mg)
was added in portions, filtered and concentrated to afford compound
mTOR, mechanistic target of rapamycin; mTORC1, mTOR
complex 1; LRS, leucylꢀtRNA synthetase; S6K1, S6 kinase 1;
MMT, monomethoxytrityl; DMF, dimethyl formamide; THF,
tetrahydrofuran; MC, methylene chloride; DCC, dicyclohexylꢀ
carbodiimide; DMAP, dimethylaminopyridine;
1
12 (0.03 mmol). Yield 56%, white solid; HꢀNMR (600MHz, CDꢀ
3OD) δ 8.52 (s, 1H, CH), 8.18 (s, 1H, CH), 6.08 (d, 1H, CH, J = 2.8
Hz), 4.64 (t, 1H, CH, J = 2.5 Hz), 4.40 (m, 1H, CH), 4.31 (m, 3H,
CH), 4.05 (d, 1H, CH, J = 1.0 Hz), 3.51 (dd, 1H, CH, J = 4.4 Hz, 1.0
Hz), 1.86 (m, 1H, CH), 1.01 (d, 3H, CH3, J = 3.2 Hz), 0.94 (d, 3H,
CH3, J = 3.2 Hz); HRMSꢀESI m/z [M+H]+ C16H24N6O9SH+ calcd
477.1325, Found: 477.1395.
■ ASSOCIATED CONTENT
Supporting Information
HPLC purities of all final compounds. Molecular formula strings.
This material is available free of charge via the Internet at
http://pubs.acs.org.
(2R,3R,4S,5R)ꢀ2ꢀ(6ꢀaminoꢀ2ꢀiodoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ5ꢀ(hydroxyl meꢀ
thyl)tetrahydrofuranꢀ3,4ꢀdiol (14). Compound 14 was prepared by
following the reported procedure.34 Yield 11% in 4 steps, yellow
1
solid; HꢀNMR (300MHz, CD3OD) δ 8.20 (s, 1H, CH), 5.89 (d, 1H,
■ REFERENCES
(1) Laplante, M.; Sabatini, D. M. mTOR signaling in growth
control and disease. Cell 2012, 149, 274ꢀ293.
(2) Ma, X. M.; Blenis, J. Molecular mechanisms of mTORꢀ
mediated translational control. Nat. Rev. Mol. Cell Biol. 2009, 10,
307ꢀ318.
(3) Dowling, R. J.; Topisirovic, I.; Alain, T.; Bidinosti, M.;
Fonseca, B. D.; Petroulakis, E.; Wang, X.; Larsson, O.; Selvaraj, A.;
Liu, Y.; Kozma, S. C.; Thomas, G.; Sonenberg, N. mTORC1ꢀ
mediated cell proliferation, but not cell growth, controlled by the 4Eꢀ
BPs. Science 2010, 328, 1172ꢀ1176.
(4) Hsieh, A. C.; Costa, M.; Zollo, O.; Davis, C.; Feldman, M. E.;
Testa, J. R.; Meyuhas, O.; Shokat, K. M.; Ruggero, D. Genetic
dissection of the oncogenic mTOR pathway reveals druggable
addiction to translational control via 4EBPꢀeIF4E. Cancer Cell 2010,
17, 249ꢀ261.
(5) Sabatini, D. M. mTOR and cancer: insights into a complex
relationship. Nat. Rev. Cancer 2006, 6, 729ꢀ734.
(6) Zoncu, R.; Efeyan, A.; Sabatini, D. M. mTOR: from growth
signal integration to cancer, diabetes and ageing. Nat. Rev. Mol. Cell
Biol. 2011, 12, 21ꢀ35.
CH, J = 6.03 Hz), 4.66 (t, 1H, CH, J = 5.31 Hz), 4.30 (dd, 1H, CH, J
= 5.10 Hz, 3.09 Hz), 4.15ꢀ4.12 (m, 1H, CH), 3.88 (dd, 1H, CH, J =
12.45 Hz, 2.73 Hz), 3.74 (dd, 1H, CH, J = 12.45 Hz, 2.94 Hz), 2.18
(s, 3H, CH3), 2.14 (s, 3H, CH3), 2.11 (s, 3H, CH3)
(2R,3R,4R,5R)ꢀ2ꢀ(6ꢀaminoꢀ2ꢀiodoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ5ꢀ(hydroxy
methyl)tetrahydrofuranꢀ3,4ꢀdiyl diacetate (15). Compound 15 was
prepared by following the procedure described for compounds 8 and
1
9. Yield 67% in 3 steps, colorless oil; HꢀNMR (300MHz, CDCl3) δ
7.77 (s, 1H, CH), 6.47 (br, 1H, OH), 5.97 (d, 1H, CH, J = 7.68 Hz),
5.91 (m, 1H, CH), 5.67 (dd, 1H, CH, J = 5.13 Hz, 1.29 Hz), 4.36 (d,
1H, CH, J = 1.08 Hz), 4.51 (dd, 1H, CH, J = 13.02 Hz, 1.47 Hz), 3.88
(d, 1H, CH, J = 11.73 Hz), 2.17 (s, 3H, CH3), 2.11 (s, 3H, CH3)
(2R,3R,4R,5R)ꢀ2ꢀ(6ꢀaminoꢀ2ꢀiodoꢀ9Hꢀpurinꢀ9ꢀyl)ꢀ5ꢀ((sulfamoyl
oxy)methyl)tetrahydrofuranꢀ3,4ꢀdiyl diacetate (16). Compound 16
was prepared by following the procedure described for compound 10.
1
Yield 86%, white solid; HꢀNMR (300MHz, CDCl3) δ 7.93 (s, 1H,
CH), 6.14 (d, 1H, CH, J = 5.49 Hz), 5.84 (br, 2H, NH2), 5.78 (t, 1H,
CH, J = 5.31 Hz), 5.71 (t, 1H, CH, J = 4.02 Hz), 4.53 (d, 2H, CH2, J =
3.48 Hz), 4.47 (m, 1H, CH), 2.16 (s, 3H, CH3), 2.09 (s, 3H, CH3)
(2R,3R,4R,5R)ꢀ2ꢀ(((Nꢀ((S)ꢀ2ꢀacetoxyꢀ4ꢀmethylpentanoyl)sulfamꢀ
oyl)oxy)methyl)ꢀ5ꢀ(6ꢀaminoꢀ2ꢀiodoꢀ9Hꢀpurinꢀ9ꢀyl)tetrahydrofuranꢀ
3,4ꢀdiyl diacetate (17). Compound 17 (0.03 mmol) was prepared by
following the procedure described for compound 11. Yield 81%,
ACS Paragon Plus Environment

Journal of Medicinal Chemistry
Page 6 of 7
(7) Chiarini, F.; Evangelisti, C.; McCubrey, J. A.; Martelli, A. M.
Moon, A.; Kim, K.; Kim, D.; Kang, E. J.; Moon, Y.; Rhee, K. H.;
Current treatment strategies for inhibiting mTOR in cancer. Trends
Pharmacol. Sci. 2015, 36, 124ꢀ135.
Han, B. W.; Yang, J. S.; Han, G.; Yang, W. S.; Lee, C.; Wang, M.
W.; Kim, S. Chemical inhibition of prometastatic lysylꢀtRNA
synthetaseꢀlaminin receptor interaction. Nat. Chem. Biol. 2014, 10,
29ꢀ34.
(23) Bharathkumar, H.; Mohan, C. D.; Rangappa, S.; Kang, T.;
Keerthy, H. K.; Fuchs, J. E.; Kwon, N. H.; Bender, A.; Kim, S.;
Basappa; Rangappa, K. S. Screening of quinoline, 1,3ꢀbenzoxazine,
and 1,3ꢀoxazineꢀbased small molecules against isolated methionylꢀ
tRNA synthetase and A549 and HCT116 cancer cells including an in
silico binding mode analysis. Org. Biomol. Chem. 2015, 13, 9381ꢀ
9387.
(24) Lynch, C. J.; Fox, H. L.; Vary, T. C.; Jefferson, L. S.;
Kimball, S. R. Regulation of amino acidꢀsensitive TOR signaling by
leucine analogues in adipocytes. J. Cell. Biochem. 2000, 77, 234ꢀ251.
(25) Wang, X.; Fonseca, B. D.; Tang, H.; Liu, R.; Elia, A.;
Clemens, M. J.; Bommer, U. A.; Proud, C. G. Reꢀevaluating the roles
of proposed modulators of mammalian target of rapamycin complex 1
(mTORC1) signaling. J. Biol. Chem. 2008, 283, 30482ꢀ30492.
(26) Yoon, S.; Kim, J. H.; Yoon, I.; Kim, C.; Kim, S. E.; Koh, Y.;
Jeong, S. J.; Lee, J.; Kim, S.; Lee, J. Discovery of (S)ꢀ4ꢀ
isobutyloxazolidinꢀ2ꢀone as a novel leucylꢀtRNA synthetase (LRS)ꢀ
targeted mTORC1 inhibitor. Bioorg. Med. Chem. Lett. 2016, 26,
3038ꢀ3041.
(27) Bernier, S.; Akochy, P. M.; Lapointe, J.; Chenevert, R.
Synthesis and aminoacylꢀtRNA synthetase inhibitory activity of
aspartyl adenylate analogs. Bioorg. Med. Chem. 2005, 13, 69ꢀ75.
(28) Forrest, A. K.; Jarvest, R. L.; Mensah, L. M.; O'Hanlon, P. J.;
Pope, A. J.; Sheppard, R. J. Aminoalkyl adenylate and aminoacyl
sulfamate intermediate analogues differing greatly in affinity for their
cognate Staphylococcus aureus aminoacyl tRNA synthetases. Bioorg.
Med. Chem. Lett. 2000, 10, 1871ꢀ1874.
1
2
3
4
5
6
7
8
(8) Hudes, G.; Carducci, M.; Tomczak, P.; Dutcher, J.; Figlin, R.;
Kapoor, A.; Staroslawska, E.; Sosman, J.; McDermott, D.; Bodrogi,
I.; Kovacevic, Z.; Lesovoy, V.; SchmidtꢀWolf, I. G.; Barbarash, O.;
Gokmen, E.; O'Toole, T.; Lustgarten, S.; Moore, L.; Motzer, R. J.;
Global, A. T. Temsirolimus, interferon alfa, or both for advanced
renalꢀcell carcinoma. N. Engl. J. Med. 2007, 356, 2271ꢀ2281.
(9) Motzer, R. J.; Escudier, B.; Oudard, S.; Hutson, T. E.; Porta,
C.; Bracarda, S.; Grunwald, V.; Thompson, J. A.; Figlin, R. A.;
Hollaender, N.; Urbanowitz, G.; Berg, W. J.; Kay, A.; Lebwohl, D.;
Ravaud, A.; Group, R.ꢀS. Efficacy of everolimus in advanced renal
cell carcinoma: a doubleꢀblind, randomised, placeboꢀcontrolled phase
III trial. Lancet 2008, 372, 449ꢀ456.
(10) Demetri, G. D.; Chawla, S. P.; RayꢀCoquard, I.; Le Cesne, A.;
Staddon, A. P.; Milhem, M. M.; Penel, N.; Riedel, R. F.; BuiꢀNguyen,
B.; Cranmer, L. D.; Reichardt, P.; Bompas, E.; Alcindor, T.; Rushing,
D.; Song, Y.; Lee, R. M.; Ebbinghaus, S.; Eid, J. E.; Loewy, J. W.;
Haluska, F. G.; Dodion, P. F.; Blay, J. Y. Results of an international
randomized phase III trial of the mammalian target of rapamycin
inhibitor ridaforolimus versus placebo to control metastatic sarcomas
in patients after benefit from prior chemotherapy. J. Clin. Oncol.
2013, 31, 2485ꢀ2492.
(11) Mita, M. M.; Gong, J.; Chawla, S. P. Ridaforolimus in
advanced or metastatic soft tissue and bone sarcomas. Expert Rev.
Clin. Pharmacol. 2013, 6, 465ꢀ482.
(12) Thoreen, C. C.; Sabatini, D. M. Rapamycin inhibits mTORC1,
but not completely. Autophagy 2009, 5, 725ꢀ726.
(13) Stipanuk, M. H. Leucine and protein synthesis: mTOR and
beyond. Nutr. Rev. 2007, 65, 122ꢀ129.
(14) Dodd, K. M.; Tee, A. R. Leucine and mTORC1: a complex
relationship. Am. J. Physiol. Endocrinol. Metab. 2012, 302, E1329ꢀ
1342.
(15) Han, J. M.; Jeong, S. J.; Park, M. C.; Kim, G.; Kwon, N. H.;
Kim, H. K.; Ha, S. H.; Ryu, S. H.; Kim, S. LeucylꢀtRNA synthetase is
an intracellular leucine sensor for the mTORC1ꢀsignaling pathway.
Cell 2012, 149, 410ꢀ424.
(16) Bonfils, G.; Jaquenoud, M.; Bontron, S.; Ostrowicz, C.;
Ungermann, C.; De Virgilio, C. LeucylꢀtRNA synthetase controls
TORC1 via the EGO complex. Mol. Cell 2012, 46, 105ꢀ110.
(17) Yao, P.; Fox, P. L. AminoacylꢀtRNA synthetases in medicine
and disease. EMBO Mol. Med. 2013, 5, 332ꢀ343.
(18) Zhao, H.; Palencia, A.; Seiradake, E.; Ghaemi, Z.; Cusack, S.;
LutheyꢀSchulten, Z.; Martinis, S. Analysis of the Resistance
Mechanism of a Benzoxaborole Inhibitor Reveals Insight into the
LeucylꢀtRNA Synthetase Editing Mechanism. ACS Chem. Biol. 2015,
10, 2277ꢀ2285.
(19) Palencia, A.; Li, X.; Bu, W.; Choi, W.; Ding, C. Z.; Easom, E.
E.; Feng, L.; Hernandez, V.; Houston, P.; Liu, L.; Meewan, M.;
Mohan, M.; Rock, F. L.; Sexton, H.; Zhang, S.; Zhou, Y.; Wan, B.;
Wang, Y.; Franzblau, S. G.; Woolhiser, L.; Gruppo, V.; Lenaerts, A.
J.; O'Malley, T.; Parish, T.; Cooper, C. B.; Waters, M. G.; Ma, Z.;
Ioerger, T. R.; Sacchettini, J. C.; Rullas, J.; AnguloꢀBarturen, I.;
PerezꢀHerran, E.; Mendoza, A.; Barros, D.; Cusack, S.; Plattner, J. J.;
Alley, M. R. Discovery of Novel Oral Protein Synthesis Inhibitors of
Mycobacterium tuberculosis That Target LeucylꢀtRNA Synthetase.
Antimicrob. Agents Chemother. 2016, 60, 6271ꢀ6280.
(20) Sonoiki, E.; Palencia, A.; Guo, D.; Ahyong, V.; Dong, C.; Li,
X.; Hernandez, V. S.; Zhang, Y. K.; Choi, W.; Gut, J.; Legac, J.;
Cooper, R.; Alley, M. R.; Freund, Y. R.; DeRisi, J.; Cusack, S.;
Rosenthal, P. J. Antimalarial Benzoxaboroles Target Plasmodium
falciparum LeucylꢀtRNA Synthetase. Antimicrob. Agents Chemother.
2016, 60, 4886ꢀ4895.
(21) Park, S. G.; Schimmel, P.; Kim, S. Aminoacyl tRNA
synthetases and their connections to disease. Proc. Natl. Acad. Sci. U.
S. A. 2008, 105, 11043ꢀ11049.
(22) Kim, D. G.; Lee, J. Y.; Kwon, N. H.; Fang, P.; Zhang, Q.;
Wang, J.; Young, N. L.; Guo, M.; Cho, H. Y.; Mushtaq, A. U.; Jeon,
Y. H.; Choi, J. W.; Han, J. M.; Kang, H. W.; Joo, J. E.; Hur, Y.;
Kang, W.; Yang, H.; Nam, D. H.; Lee, M. S.; Lee, J. W.; Kim, E. S.;
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
(29) Lee, J.; Kim, S. E.; Lee, J. Y.; Kim, S. Y.; Kang, S. U.; Seo, S.
H.; Chun, M. W.; Kang, T.; Choi, S. Y.; Kim, H. O. Nꢀ
Alkoxysulfamide, Nꢀhydroxysulfamide, and sulfamate analogues of
methionyl and isoleucyl adenylates as inhibitors of methionylꢀtRNA
and isoleucylꢀtRNA synthetases. Bioorg. Med. Chem. Lett. 2003, 13,
1087ꢀ1092.
(30) Kim, S. E.; Kim, S. Y.; Kim, S.; Kang, T.; Lee, J.
Deoxyribosyl analogues of methionyl and isoleucyl sulfamate
adenylates as inhibitors of methionylꢀtRNA and isoleucylꢀtRNA
synthetases. Bioorg. Med. Chem. Lett. 2005, 15, 3389ꢀ3393.
(31) Gadakh, B.; Smaers, S.; Rozenski, J.; Froeyen, M.; Van
Aerschot,
A.
5'ꢀ(Nꢀaminoacyl)ꢀsulfonamidoꢀ5'ꢀdeoxyadenosine:
attempts for a stable alternative for aminoacylꢀsulfamoyl adenosines
as aaRS inhibitors. Eur. J. Med. Chem. 2015, 93, 227ꢀ236.
(32) Lukkarila, J. L.; da Silva, S. R.; Ali, M.; Shahani, V. M.; Xu,
G. W.; Berman, J.; Roughton, A.; DheꢀPaganon, S.; Schimmer, A. D.;
Gunning, P. T. Identification of NAE Inhibitors Exhibiting Potent
Activity in Leukemia Cells: Exploring the Structural Determinants of
NAE Specificity. ACS Med. Chem. Lett. 2011, 2, 577ꢀ582.
(33) Filippov, D.; Timmers, C. M.; Roerdink, A. R.; van der Marel,
G. A.; van Boom, J. H. Synthesis of the antibiotically active part of
agrocin 84. Tetrahedron Lett. 1998, 39, 4891ꢀ4894.
(34) Piton, N.; Mu, Y.; Stock, G.; Prisner, T. F.; Schiemann, O.;
Engels, J. W. Baseꢀspecific spinꢀlabeling of RNA for structure
determination. Nucleic Acids Res. 2007, 35, 3128ꢀ3143.
(35) Vichai, V.; Kirtikara, K. Sulforhodamine B colorimetric assay
for cytotoxicity screening. Nat. Protoc. 2006, 1, 1112ꢀ1116.
(36) Wang, X.ꢀW.; Zhang, Y.ꢀJ. Targeting mTOR network in
colorectal cancer therapy. World J. Gastroenterol. 2014, 20, 4178.
(37) Francipane, M. G.; Lagasse, E. mTOR pathway in colorectal
cancer: an update. Oncotarget 2014, 5, 49ꢀ66.
(38) Milton, S.; Ander, C.; Honcharenko, D.; Honcharenko, M.;
Yeheskiely, E.; Strömberg, R. Synthesis and Stability of a 2′
[N (Aminoethyl) carbamoyl] methyladenosine Containing
Dinucleotide. Eur. J. Org. Chem. 2013, 2013, 7184ꢀ7192.
- O-
-
-
ACS Paragon Plus Environment

Page 7 of 7
Journal of Medicinal Chemistry
Authors are required to submit a graphic entry for the Table of Contents (TOC) that, in conjunction with the manuscript title,
should give the reader a representative idea of one of the following: A key structure, reaction, equation, concept, or theorem,
etc., that is discussed in the manuscript. Consult the journal’s Instructions for Authors for TOC graphic specifications.
1
2
3
4
5
Table of Contents
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
7
ACS Paragon Plus Environment