2-Oxo-N-aryl-1,2,3,4-tetrahydroquinoline-6-sulfonamides as activators of the tumor cell specific M2 isoform of pyruvate kinase

Article abstract of DOI:10.1016/j.bmcl.2011.08.114

Compared to normal differentiated cells, cancer cells have altered metabolic regulation to support biosynthesis and the expression of the M2 isozyme of pyruvate kinase (PKM2) plays an important role in this anabolic metabolism. While the M1 isoform is a highly active enzyme, the alternatively spliced M2 variant is considerably less active and expressed in tumors. While the exact mechanism by which decreased pyruvate kinase activity contributes to anabolic metabolism remains unclear, it is hypothesized that activation of PKM2 to levels seen with PKM1 may promote a metabolic program that is not conducive to cell proliferation. Here we report the third chemotype in a series of PKM2 activators based on the 2-oxo-Naryl-1,2,3,4-tetrahydroquinoline-6-sulfonamide scaffold. The synthesis, structure activity relationships, selectivity and notable physiochemical properties are described.

Full text of DOI:10.1016/j.bmcl.2011.08.114

Bioorganic & Medicinal Chemistry Letters 21 (2011) 6322–6327
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
2-Oxo-N-aryl-1,2,3,4-tetrahydroquinoline-6-sulfonamides as activators
of the tumor cell specific M2 isoform of pyruvate kinase
Martin J. Walsh a, Kyle R. Brimacombe a, Henrike Veith a, James M. Bougie a, Thomas Daniel a,
William Leister a, Lewis C. Cantley b,c, William J. Israelsen d, Matthew G. Vander Heiden d,e
,
Min Shen a, Douglas S. Auld a, , Craig J. Thomas a, Matthew B. Boxer a,
⇑
a NIH Chemical Genomics Center, NIH Center for Translational Therapeutics, National Human Genome Research Institute, National Institutes of Health, 9800 Medical Center
Drive, Rockville, MD 20850, USA
b Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, MA 02115, USA
c Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA
d Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
e Dana-Farber Cancer Institute, Boston, MA 02115, USA
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 14 July 2011
Revised 25 August 2011
Accepted 26 August 2011
Available online 14 September 2011
Compared to normal differentiated cells, cancer cells have altered metabolic regulation to support bio-
synthesis and the expression of the M2 isozyme of pyruvate kinase (PKM2) plays an important role in this
anabolic metabolism. While the M1 isoform is a highly active enzyme, the alternatively spliced M2 var-
iant is considerably less active and expressed in tumors. While the exact mechanism by which decreased
pyruvate kinase activity contributes to anabolic metabolism remains unclear, it is hypothesized that acti-
vation of PKM2 to levels seen with PKM1 may promote a metabolic program that is not conducive to cell
proliferation. Here we report the third chemotype in a series of PKM2 activators based on the 2-oxo-N-
aryl-1,2,3,4-tetrahydroquinoline-6-sulfonamide scaffold. The synthesis, structure activity relationships,
selectivity and notable physiochemical properties are described.
Keywords:
PKM2
Pyruvate kinase
Cellular metabolism
Anti-cancer strategies
Small molecule activators
Published by Elsevier Ltd.
The M2 isoform of pyruvate kinase (PKM2) is expressed in
tumor cells and has been proposed as a diagnostic marker for a
large number of cancers.1 While the expression of this isoform in
tumors has been known for a number of years,2 recent work3–5
has invigorated considerable interest in understanding pyruvate
kinase regulation and the possibility of targeting PKM2 for cancer
therapy.6–8 One study has shown that the aberrant metabolism
of cancer cells and their production of high levels of lactate in aer-
obic conditions could be reversed by replacement of the M2 iso-
form of pyruvate kinase (PK) with the M1 isoform.3 Rescue of
cells subjected to shRNA knock-down of PKM2 with PKM1 also
showed reduced tumor formation in a xenograft model. Pyruvate
kinase functions in glycolysis to catalyze conversion of phospho-
enolpyruvate (PEP) to pyruvate with concomitant generation of
ATP from ADP. There are four mammalian PK isoforms encoded
by two genes located on two distinct chromosomes.9–13 Encoded
by PKLR, PKR is found in erythrocytes and PKL is expressed in both
the liver and kidney. The PKM gene also has two splice varients;
PKM1 that is predominantly found in muscle and brain tissue
and PKM2 that is expressed in some adult tissues, embryonic tis-
sues and tumor cells.1 Regardless of the isoform, the active form
of PK is a tetramer. PKM2, PKL and PKR are allosterically activated
in a feed forward mechanism by fructose-1,6-bisphosphate (FBP)
while PKM1, which is expressed in tissues with high ATP require-
ments, does not require allosteric activation.14 While it has been
reported that PKM2 in tumors exists predominantly as an inactive
dimer,1,15 exactly how this contributes to tumorigenesis and the
Warburg effect is still unknown.
It has been hypothesized that cancer cells benefit from the low
activity of PKM2 to increase levels of glycolytic intermediates for
use in anabolic processes including nucleic acid, amino acid, and li-
pid syntheses.15 This suggests that pharmacological activation of
PKM2 to levels associated with PKM1 may inhibit cell proliferation
as well as be a potential therapeutic strategy for cancer. Our group
has previously reported two chemotypes capable of activating
PKM2 including a series of diarylsulfonamides16 and a series of
thieno[3,2-b]pyrrole[3,2-d]pyridazinones17 (exemplified by 1 and
2, respectively, in Fig. 1). Here, we describe the development of a
series of PKM2 activators derived from the 2-oxo-N-aryl-tetrahy-
droquinoline-6-sulfonamide lead, 3.
⇑
Corresponding author. Tel.: +1 301 217 4681; fax: +1 301 217 5736.
E-mail address: boxerm@mail.nih.gov (M.B. Boxer).
Present address: Novartis Institutes for Biomedical Research, Cambridge,
MA 02139, USA
0960-894X/$ - see front matter Published by Elsevier Ltd.
doi:10.1016/j.bmcl.2011.08.114

M. J. Walsh et al. / Bioorg. Med. Chem. Lett. 21 (2011) 6322–6327
6323
O
O
S
O
O
S
O
Me
O
N
O
N
N
S
NH
N
N
O
OMe
Me
NH
O
S
S
O
NH2
O
NCGC00185916 (1)
hpkM2 AC50: 38 nM
NCGC00186527 (2)
hpkM2 AC50: 73 nM
NCGC00185939 (3)
qHTS hit
hpkM2 AC50: 790 nM
Figure 1. Chemical structures of previously reported hPKM2 activators NCGC00185916 (1), NCGC00186527 (2) and qHTS Hit NCGC00185939 (3).
Table 1
Table 2
SAR of N-(3,4-dimethylphenyl)arylsulfonamides
SAR of 2-oxo-1,2,3,4-tetrahydroquinoline-6-sulfonamides
R
O
O
S
O
Me
O
HN
R
NH
S
O
Me
NH
a
#
R
hPK M2 AC50
M)
hPK M2 Max
Resp.b (%)
a
#
R
hPK M2 AC50 hPK M2 max
(
l
(lM)
resp.b (%)
23 2-Methyl-benzene
24 3-Methyl-benzene
25 4-Methyl-benzene
26 2-Fluoro-benzene
27 3-Fluoro-benzene
28 4-Fluoro-benzene
29 3-Chloro-benzene
30 4-Chloro-benzene
31 4-(Pyridine)
2.2
0.25
1.4
5.2
0.79
6.0
0.41
2.0
>20
>20
16.3
1.1
94
4
5
6
7
8
9
5-(N-Methylindole)
3-(Benzoic acid)
4-Fluoro-C6H4
2-Napthalene
3-Chloro-C6H4
19.9
>20
>20
18.4
18.3
17.3
10.0
47
NA
NA
69
71
53
62
81
NA
58
76
57
88
101
102
95
101
84
104
106
NA
NA
85
115
NA
100
4-Methoxy-C6H4
10 6-(2,2-Dimethylchroman)
11 6-(2H-Benzo[b][1,4]oxazin-3(4H)-one) 13.4
12 5-(1H-Benzo[d]imidazol-2(3H)-one)
13 3-NHAcetyl-C6H4
14 6-(N-Acetyl-tetrahydroquinoline)
15 5-(Indolin-2-one)
16 7-(4,5-Dihydro-1H-benzo[b]azepin-
2(3H)-one)
17 4-NHAcetyl-C6H4
18 6-(3-Methylbenzo[d]oxazol-2(3H)-
one)
19 6-(4-Methyl-3,4-dihydro-2H-
benzo[b][1,4]oxazine)
>20
13.1
9.0
8.3
7.1
32 3-(Pyridine)
33 4-Methoxy-benzene
34 3-Methoxy-benzene
35 3-Phenyl-benzene
36 6-(2,3-
Dihydrobenzo[b][1,4]dioxine)
37 3-Trifluoromethyl-benzene
38 2-Napthalene
>20
11.1
6.9
4.3
<40
95
6.4
1.3
>20
4.6
0.10
1.3
0.56
0.54
0.29
0.44
109
121
NA
73
104
95
121
138
123
128
39 6-(Quinoline)
6.0
81
40 5-(2,3-Dihydro-1H-indene)
41 3-Chloro-4-methyl-benzene
42 3,4-Dichloro-benzene
43 4-Chloro-3-methyl-benzene
44 3-Fluoro-4-methylbenzene
45 3-Chloro-4-fluoro-benzene
46 4-Fluoro-3-methyl-benzene
20 4-(2-Oxopyrrolidin-1 -yl)-C6H4
21 5-(1-Methylindolin-2-one)
22 5-(N-Acetyllindoline)
0.94
2.3
1.5
60
119
96
a
AC50 values were determined utilizing the luminescent pyruvate kinase–lucif-
erase coupled assay (Ref. 16) and the data represents the average from three sep-
arate experiments.
Max Res. (Maximum Response) is % activity that represents the % activation at
57 nM of compound. See Supplementary data for normalization.
a
AC50 values were determined utilizing the luminescent pyruvate kinase–lucif-
erase coupled assay (Ref. 16) and the data represents the average from three sep-
arate experiments.
b
b
Max Res. (Maximum Response) is % activity that represents the % activation at
57
l
M of compound. See Supplementary data for normalization.
Using our previously described qHTS assay for PKM2 activa-
tors,16 compound 3 was found to have an AC50 = 790 nM in the pri-
mary screen. Resynthesis of the hit as well as the majority of
analogs described in Tables 1 and 2 was accomplished via base-
mediated coupling of commercially available anilines and sulfonyl
chlorides in DMF, as represented in Scheme 1. An array of commer-
cially available anilines were coupled with sulfonyl chlorides using
either Hunig’s base or pyridine in DMF solvent.
R2
Hunig's Base
NH2
R2
or
O
pyridine
O
+
O S
S
O
DMF, rt, 1 h
NH
Cl
R1
R1
We began by looking at potential modification of the aryl link-
age, but the importance of the NH-sulfonamide was apparent after
both the N-methylsulfonamide and amide-linked derivatives
showed a complete loss in activity (data not shown). We next
investigated the importance of the 3,4-dihydroquinolin-2(1H)-
one moiety by removing of the amide functionality to generate a
number of simple substituted benzene derivates (5–9). While a
number of these retained activity, they all showed significantly
diminished potency (P20-fold) and/or efficacy. Informed by these
results, we next investigated various amide, urea and carbamate
derivatives as well as heteroatom inclusion. A few interesting
SAR points from these modifications were: placing the amide car-
bonyl outside of the ring (14 and 22), changing the amide ring
Scheme 1. General synthetic methodology used to access analogs in Tables 1 and 2.
structure to a five- or seven-membered rings (15 and 16) or remov-
ing the cyclic nature of the amide (13 and 17), all resulted in a sig-
nificant loss of potency. In general, most analogs that altered the
six-membered lactam moiety saw a considerable drop in activity.
The next SAR we wanted to explore was altering the
3,4-dimethylaniline moiety by addition of a variety of substituted
anilines to the commercially available 2-oxo-1,2,3,4-tetrahydro-
quinoline-6-sulfonyl chloride (Table 2). Preference for substitution
of hydrophobic groups at the meta-position quickly emerged as a
key for potency as the 3-methyl (24), 3-fluoro (27) and 3-chloro

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M. J. Walsh et al. / Bioorg. Med. Chem. Lett. 21 (2011) 6322–6327
Table 3
SAR of 3,4-dihydroquinolin-2(1H)-one-7-sulfonamides, 3,4-dihydroquinolin-2(1H)-one-6-sulfonamides and 3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-7-sulfonamides
a
#
R1
R2
R3
hPK M2 AC50
(l
M)
hPK M2 Max Resp.b (%)
47
48
Phenyl
Me
Me
Me
Me
>20
>20
NA
NA
H
N
R1
O
R2
R3
Ph
Methyl
CI
F
O
49
50
51
Me
Me
Me
Me
Me
Me
0.48
0.26
0.21
111
80
101
S
X
N
H
O
47-50 X = O
52
53
54
Me
Me
Me
Me
Me
Me
0.22
3.8
129
106
102
Me2N
Et2N
MeHN
N
51-70, X = CH2
0.44
55
56
Me
Me
Me
Me
0.51
>20
123
NA
N
Me2HN
N
57
58
Me
Me
Me
Me
18.3
0.57
47
123
N
H
N
H
59
60
Me
Me
Me
Me
2.1
83
HO
HO
N
H
0.18
132
HO
HO
HO
61
62
63
Me
Me
Me
Me
Me
Me
0.23
0.88
0.17
124
108
116
N
H
N
H
N
H
64
65
66
67
F
F
F
F
Me
Cl
F
Cl
Me
Cl
0.14
0.60
0.09
0.24
112
93
88
F
Me
92
HO
HO
HO
68
69
70
Me
F
Cl
.06
96
92
95
N
H
Cl
0.13
0.29
N
H
F
Me
N
H
R1
O
71
72
Cl
Cl
Me
Me
Me
Cl
0.07
0.09
92
109
R2
R3
73
Me
Me
0.21
106
Me2N
Br
Br
S
N
H
O
N
74a
75a
Me
Me
Me
Cl
6.60
7.43
52
51
O
H
71-77
HO
HO
76a
77a
Me
Me
Me
Cl
6.1
6.1
68
54
N
H
N
H
a
AC50 values were determined utilizing the luminescent pyruvate kinase–luciferase coupled assay (Ref. 16) and the data represents the average from three separate experiments.
Max Res. (Maximum Response) is % activity that represents the % activation at 57 mM of compound. See Supplementary data for normalization.
b
(29) analogs all had AC50 values <800 nM. Notably, the 3-methoxy
analog (34) saw a potency drop of >4-fold compared to the
marked decrease in activity while more tolerance was observed
at the para-position. Noteworthy inactive analogs were pyridine
derivatives 31 and 32 as well as the quinoline derivative (39) for
3-methyl version. All ortho-substituents tested resulted in
a

M. J. Walsh et al. / Bioorg. Med. Chem. Lett. 21 (2011) 6322–6327
6325
A
H
H
N
H
N
Br
O
O S
N
O
Ar
O
O S
O
NH2
Br
O
Ar-B(OH)2, tetrakis (3 mol%)
Hunig's Base
DMF, rt, 1 h
O
S
+
Me
O
O
O
Me
Me
NH
Me
Me
NH
Na2CO3, 1,2,-DME/H2O
µW, 120 °C, 20 min
Cl
O
Me
B
H
R2
N
H
F
O
O S
N
O
H
N
NH2
H
N
F
S
N
O
O
R
R3
R2
Hunig's Base
DMF, rt, 1 h
3 O
O S
NH
O
+
NH
MeCN, µW, 180 °C
Cl
R1
O
R1
R1
Scheme 2. General synthetic methodology used to access analogs in Table 3.
which the non-nitrogenous 2-naphthalene analog (38) had reason-
able activity (1.3 M). Tethering the hydrophobic dimethyl group
Good activity was obtained starting with 7-fluoro-2-oxo-
1,2,3,4-tetrahydroquinoline-6-sulfonyl chloride to generate 51
with a potency of 210 nM. This particular analog was further mod-
ified by introduction of nitrogen nucleophiles through SNAr reac-
tions in acetonitrile (Scheme 2B). Significant SAR was seen with
substitution of a variety of these analogs. Sterically larger groups
like N,N-dimethylpyrrolidin-3-amine (56) and piperidine (57) re-
sulted loss of activity while smaller dimethyl- (52) and
methyl-amine (54) derivatives retained activity. With the realiza-
tion that small amines were tolerated at this position a few analogs
were generated in an effort to improve both solubility and potency
(58–63). The racemic alaninol derivative provided high aqueous ki-
l
in a dihydroindene ring resulted in a much less potent analog
(40), while combinations of methyl, fluoro and chloro at the
3- and 4-positions resulted in a number of analogs with good
potencies (41–46). Both the 3-chloro-4-methyl (41) and
3-chloro-4-fluoro (45) derivatives had potencies <300 nM, so
these, along with the original 3,4-dimethyl substitution patterns
were chosen as motifs to carry on.
Derivatives with various substituents at the 6-position of 3,4-
dihydroquinolin-2(1H)-one-7-sulfonamides, the 7-position of
3,4-dihydroquinolin-2(1H)-one-6-sulfonamides, and the 6-position
of
3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazine-7-sulfonamides
netic solubility (>120 lM in PBS buffer) while retaining low nano-
were examined next (Table 3). 6-bromo-3-oxo-3,4-dihydro-2H
-benzo[b][1,4]oxazine-7-sulfonyl chloride was coupled with 3,4-
dimethylaniline and then subjected to standard Suzuki cross
coupling conditions to provide a variety of aryl substituted dihy-
dro-2H-benzo[b][1,4]oxazines (Scheme 2A). Generally, the addition
of aryl and heteroaryl substituents resulted in decreased reactivity
(exemplified by 47 and 48 in Table 3).
molar potency (61). Preparation of the enantiopure versions with
(S)- and (R)-alaninol resulted the (S)-derivative being four-fold
more potent than its enantiomer (880 nM for 62 vs 170 nM for
63). Combination of 7-fluoro or 7-(S)-alaninol derivatives with
the best aniline analogs from Table 2 gave compounds 64–70,
which all have low nanomolar activity with good maximum
response. Interestingly, it was discovered that flipping the amide
Br
Br
Br
H
N
1) ClSO3H, 65 °C
H3C
S
N
H
O
N
H
O
O O
N
H
O
2) 79 or 80, Et3N
S
O
O
H3C
R
N
H
THF, rt
R
Yield = 40–50%
~7:1 a/b
78
74a
75a
74b
75b
R = CH3
R = Cl
R = CH3
R = Cl
NH2
NH2
CuI,
Pd2(dba)3,
t-BuXPhos, NaOtBu
Toluene, 100 °C
CH3
CH3
HO
HO
1,10-phenanthroline
Cs2CO3
H3C
H3C
Cl
NH2
NH2
CH3
79
DMSO, 150 °C
80
CH3
CH3
HO
HO
HN
HN
H
N
H3C
S
N
H
O
N
H
O
O O
S
N
O
O
H3C
R
H
R
76a
77a
76b
77b
R = CH3
R = Cl
R = CH3
R = Cl
Scheme 3. Synthesis of 6-substituted 3,4-dihydroquinolin-2(1H)-one-7-sulfonamides

6326
M. J. Walsh et al. / Bioorg. Med. Chem. Lett. 21 (2011) 6322–6327
Figure 2. Pyruvate kinase isoform selectivity profile.
orientation to generate the 6-chloro- and 6-(dimethylamino)-3,4-
dihydroquinolin-2(1H)-one-7-sulfonamide derivatives gave potent
analogs 71–73. These results led us to develop chemistry to quickly
access these constitutional isomers (Scheme 3). The sulfonylation
of 78 generated regioisomers that after coupling with 79 afforded
74a and 74b or with aniline 80 afforded 75a and 75b. Analogs
significant decrease in Km for PEP (Km ꢀ1.9 mM without 66,
0.4 mM with 66) while not affecting the Km of ADP (Km ꢀ0.1 mM
in both conditions). A number of analogs were also taken on to test
their cell permeability and in vitro microsomal stability in both
mouse and human liver microsomes (Caco-2, MLM and HLM in Ta-
ble 4). While the original lead (3) had moderate Caco-2 permeabil-
ity with an efflux ratio <1, it had poor microsomal stability in both
mouse and human liver microsomes (t1/2 <20 min). It became
apparent that while the alaninol substituent provided good solu-
bility and potency, high efflux ratios and reduced MLM half lives
were attributable to the incorporation of this side-chain (68 and
69 in Table 4). A more drastic reduction in microsomal stability
was seen with the N-methyl- and N,N-dimethyl-derivatives 52
and 54 both having half lives <25 min. This data suggests that al-
kyl-amine substitution at this position engenders the molecules
with efflux and/or microsomal stability liabilities. Compounds
showing good permeability and efflux ratios (61) had either
hydrogen or fluorine substituents at the 7-position (3, 64 and
66). The metabolic liabilities of the meta- and para-groups was as-
sessed by comparing analogs 51, 64 and 66. Switching the meta-
methyl group to a meta-chlorine slightly decreased both human
and mouse microsomal stability (compare 64 to 51). Further,
74a and 75a had weak activity (>5 lM) while their regioisomers,
74b and 75b had no activity (data not shown). The incorporation
of the (S)-alaninol through Ullmann chemistry for the active bro-
mide isomers generated 76a and 77a, but rather surprisingly, these
compounds had significantly diminished activity compared to ana-
logs 63 and 68, respectively. Buchwald chemistry was required for
the conversion of 74b and 75b to the alaninol-substituted deriva-
tives 76b and 77b, respectively. Both of these analogs proved to
be inactive, as well (data not shown).
Having generated a number of analogs with potencies <200 nM,
a select few were taken on to assess their selectivity against the
other PK isoforms (Fig. 2). All analogs tested had no activity against
the R or M1 isoforms and showed only minimal activation of the
L
isoform (625% at 57.5 lM). Compound 66 was also tested for
its ability to modulate PEP and ADP affinity for hPKM2 (Fig. 3).
As with our previously reported activators,16,17 this analog showed
Figure 3. Compound 66 affects PEP affinity, but not ADP affinity. (A) Initial velocity as a function of ADP concentration in the absence (s) or presence (d) of 66 (10
lM). (B)
Initial velocity as a function PEP concentration in the absence (s) or presence (d) of 66 (10
lM). Vo, initial rate in pmol/min as determined in the PK-LDH coupled assay
(kinetic assays were carried out at approximately 22 oC with 10 nM PKM2 using [KCl] = 200 mM, [MgCl2] = 15 mM, and with either [ADP] or [PEP] = 4.0 mM; see
Supplementary data).

M. J. Walsh et al. / Bioorg. Med. Chem. Lett. 21 (2011) 6322–6327
6327
Table 4
Microsomal stability and caco-2 permeability of select compounds
Supplementary data
Compound
In vitro
Caco-2 permeabilitya
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.bmcl.2011.08.114.
microsoaml;
stabilitya t1/2
(min)
Papp (A–B)
(10À6, cm/
s)
Papp (B–A)
Efflux
ratio
(10À6, cm/
s)
References and notes
HLM
MLM
3
6.0
17.3
54.1
117.5
36.3
22.9
21.7
34.3
19.1
14.9
6.22
9.38
1.61
1.13
ND
12.21
6.24
7.91
17.52
11.9
ND
0.82
1
0.84
10.91
10.51
ND
1. Mazurek, S.; Boschek, C. B.; Hugo, F.; Eigenbrodt, E. Semin. Cancer Biol. 2005, 15,
300.
2. Reinacher, M.; Eigenbrodt, E. Virchows Arch. B. Cell Pathol. Incl. Mol. Pathol. 1981,
37, 79.
3. Christofk, H. R.; Vander Heiden, M. G.; Harris, M. H.; Ramanathan, A.; Gerszten,
R. E.; Wei, R.; Fleming, M. D.; Schreiber, S. L.; Cantley, L. C. Nature 2008, 452,
230.
4. Christofk, H. R.; Vander Heiden, M. G.; Wu, N.; Asara, J. M.; Cantley, L. C. Nature
2008, 452, 181.
64
66
69
68
52
51
54
22.9
277.2
78.8
30.4
19.4
16.5
19.7
ND
ND
ND
ND
ND
ND
a
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See Supplementary data for experimental procedures.
switching the para-methyl group on 64 to a para-fluorine on 66
saw a marked increase in both HLM and MLM with a half life of
277.2 and 117.5 min, respectively.
In conclusion, we have developed a third novel chemotype
capable of activating PKM2. These molecules, based on the 2-
oxo-N-aryl-1,2,3,4-tetrahydroquinoline-6-sulfonamide
scaffold,
have low nanomolar potency with a notable analog, 66, having
an AC50 of 90 nM with selectivity over other PK isoforms, good
Caco-2 permeability, a low efflux ratio (0.84) and high microsomal
stability (t1/2 = 277.2 min in HLM and 117.5 min in MLM).
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Acknowledgments
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We thank Paul Shinn, Danielle VanLeer and Christopher LeClair
for assistance with compound management. This research was
supported by the Molecular Libraries Initiative of the NIH Roadmap
for Medical Research and the Intramural Research Program of the
National Human Genome Research Institute, National Institutes
of Health.