Discovery of N-Cyano-sulfoximineurea Derivatives as Potent and Orally Bioavailable NLRP3 Inflammasome Inhibitors

Article abstract of DOI:10.1021/acsmedchemlett.9b00433

NLRP3 inflammasome mediated release of interleukin-1β (IL-1β) has been implicated in various diseases. In this study, rationally designed mimics of sulfonylurea moiety were investigated as NLRP3 inhibitors. Our results culminated into discovery of series of unprecedented N-cyano sulfoximineurea derivatives as potent NLRP3 inflammasome inhibitors. Compound 15 (IC₅₀ = 7 nM) and analogues were found to be highly potent and selective NLRP3 inflammasome inhibitor with good pharmacokinetic profile. These effects translate in vivo, as 15, 29, and 34 significantly inhibit NLRP3 dependent IL-1β secretion in mice.

Full text of DOI:10.1021/acsmedchemlett.9b00433

Subscriber access provided by Karolinska Institutet, University Library
Letter
Discovery of N-Cyano-Sulfoximineurea Derivatives as Potent
and Orally Bioavailable NLRP3 Inflammasome Inhibitors
Sameer Agarwal, Santosh Sasane, Hardik A. Shah, Jignesh P. Pethani, Prashant Deshmukh,
Vismit Vyas, Pravin Iyer, Harsh Bhavsar, Kasinath Viswanathan, Debdutta Bandyopadhyay,
Poonam Giri, Jogeswar Mahapatra, Abhijit Chatterjee, Mukul R Jain, and Rajiv Sharma
ACS Med. Chem. Lett., Just Accepted Manuscript • DOI: 10.1021/acsmedchemlett.9b00433 • Publication Date (Web): 27 Feb 2020
Downloaded from pubs.acs.org on February 28, 2020
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 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 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.
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 11
ACS Medicinal Chemistry Letters
1
2
3
4
5
6
7
8
9
Discovery of N-Cyano-Sulfoximineurea Derivatives as Potent
and Orally Bioavailable NLRP3 Inflammasome 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
Sameer Agarwal*, Santosh Sasane, Hardik A. Shah, Jignesh P. Pethani, Prashant Deshmukh,
Vismit Vyas, Pravin Iyer, Harsh Bhavsar, Kasinath Viswanathan, Debdutta Bandyopadhyay,
Poonam Giri, Jogeswar Mahapatra, Abhijit Chatterjee, Mukul R. Jain, and Rajiv Sharma*
Zydus Research Centre, Cadila Healthcare Ltd., Sarkhej-Bavla N.H. No. 8 A, Moraiya, Ahmedabad – 382 210, India.
KEYWORDS: NLRP3, NLRP3 inflammasome, Interleukin-1β (IL-1β), Inflammation, Sulfonylurea, N-cyano
sulfoximineurea derivatives.
ABSTRACT: NLRP3 inflammasome mediated release of interleukin-1β (IL-
1β) has been implicated in various diseases. In this study, rationally designed
mimics of sulfonylurea moiety were investigated as NLRP3 inhibitors. Our
results culminated into discovery of series of unprecedented N-cyano
sulfoximineurea derivatives as potent NLRP3 inflammasome inhibitors.
Compound 15 (IC50 = 7 nM) and analogs were found to be highly potent and
selective NLRP3 inflammasome inhibitor with good pharmacokinetic
profile. These effects translate in vivo, as 15, 29 and 34 significantly inhibit
NLRP3 dependent IL-1β secretion in mice.
Inflammasomes are multi-protein complexes formed by
innate immune sensors, including nucleotide
bowel disease (IBD), juvenile arthritis, neurodegenerative
and autoimmune diseases.6,7 Hence, the NLPR3
inflammasome might be a potential target for the
treatment of these complex diseases.8,9
oligomerization domain (NOD)-like receptor protein
(NLR) family members NLRP1, NLRP3, and NLRC4, along
with other non-NLR receptors, such as AIM2 and IFI16.1
NLRP3 inflammasome activation is dependent on two
successive signals. The first step comprises a initiating
signal (priming) in which many danger associated
molecular patterns (DAMPs) and pathogen associated
molecular patterns (PAMPs) are recognized by TLRs,
Additionally, advantage of blocking NLRP3 over simply
using an immunosuppressant is that it targets this
inflammatory response while leaving the rest of the
immune system to operate as normal.10 Therefore, NLRP3
is emerging as a promising target to develop novel and
specific compounds for treatment of anti-inflammatory
and autoimmune diseases.11 Current treatments for these
diseases include biologic drugs that target IL-1β, such as
anakinra, canakinumab, and rilonacept.12 However, they
also have major limitations such as inconvenient
treatment routes and high cost. Hence, an oral small
molecule inhibitor of NLRP3 is desirable. This has
attracted a great deal of attention in the pharmaceutical
industry and in academia.13-15
which
in
turn
up-regulates
transcription
of
inflammasome-related components, including inactive
NLRP3, proIL-1β, and proIL- 18.2,3 In the second step of
inflammasome activation, the oligomerization of NLRP3
and subsequent assembly of NLRP3, ASC, and procaspase-
1 results in formation of a complex. This triggers the
transformation of procaspase-1 to caspase-1, leading to the
release of active pro-inflammatory cytokines IL-1β and IL-
18.4
Several structurally diverse small molecule modulators of
NLRP3 have been described (Figure 1). MCC950 is
reported to be a small molecule inhibitor of NLRP3
inflammasome with an early promise for treatment of
inflammatory diseases.16 In addition, novel boron
compound (NBC-6),17 sulfonamide (JC-171)18 and
compounds like CY0914 and OLT117719 have also been
reported as NLRP3 inflammasome inhibitors. Oridonin20
and Tranilast21 have recently also been reported to block
the NLRP3 inflammasome pathways.
The NLRP3 inflammasome is the most well understood
and widely studied owing to its role in host defense and
innate immunity.5 The NLRP3 inflammasome is key
component of the inflammatory response. The
inappropriate activation of NLRP3 is implicated in a wide
range of diseases including rare periodic fever syndrome,
CAPS, TRAPS and variety of human complex diseases
such as multiple sclerosis, atherosclerosis, alzheimer’s
disease, diabetes, asthma, gouty arthritis, inflammatory
ACS Paragon Plus Environment

ACS Medicinal Chemistry Letters
Page 2 of 11
all these performed modulation lowered the activity
obtaining less than 50% inhibition of IL-1β release at 1 µM.
1
2
3
4
5
6
7
O
NH2
O
B
O
O
Table 1. Core modification on the sulfonylurea moiety: In
vitro activity of MCC950 (1) and 2-20.a
O
HO
Cl3C
S
N
N
SO2NHOH
H
H
O
MCC-950
NBC-6
R
X
8
9
S
O
O
S
O
NH
N
S
CN
MeO
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
OLT-1177
O
O
CF3
Cl
CY-09
JC-171
OH
Figure 1. Selected known NLRP3 Inflammasome Inhibitors.
This communication describes the discovery of a series of
unprecedented N-cyano sulfoximineurea derivatives as
highly potent, orally bioavailable, and selective NLRP3
inflammasome inhibitors. We hypothesized that
rationally designed mimics of sulfonylurea moiety in the
MCC950 would result in structurally novel NLRP3
inflammasome inhibitors with improved properties
(Figure 2). Accordingly, we thus focused our SAR efforts
on exploring this region of the molecule.
O
O
O
HO
S
R
N
N
X
H
H
O
MCC-950
Figure 2. Schematic representation of ligand optimization.
A series of novel compounds have been synthesized by
rational and systematic bioisosteric replacement of
sulfonylurea (Figure 2). Drug-like properties such as
molecular weight (<500) and clog P of these NCEs are
generally within acceptable range of oral drugs.22 The
ability of test compounds to inhibit NLRP3 inflammasome
was measured in IL-1β assay using THP1 cells.16 Table 1
describes the structure-activity relationship (SAR) of
replacement of sulfonylurea group. The most studied
NLRP3 inhibitor, MCC950 (also called CRID3 and
CP-456,773) was found to be potent in our hands as well
(IC50 = 8 nM); and its corresponding phenyl (2) and tosyl
(3) analogues were also potent with IC50 of 35 nM and 15
nM respectively. Introduction of cyanomethyl urea or 1-
(2,2,2-trifluoroethyl) urea in place of sulfonylurea, (as
exemplified by analogs 4 and 5) resulted in loss of activity
with less than 50% inhibition of IL-1β release at 1 µM.
Changing the topography by introducing oxetane or
squaric acid moiety also resulted in dramatic potency loss
(6 and 7). Further, various other rationally designed
mimics of sulfonylurea moiety like 2-oxo-N-
(arylsulfonyl)acetamide derivative 8, corresponding
hydroxy derivative 9 and acetamide 10 also lowered the
activity. Moreover, sulfonamide derivative 11, oxalamide
derivative 12, phosphonamidic acid analogue 13, and
cyanoguanidine derivative 14 were also synthesized and
explored as NLRP3 inflammasome inhibitors. However,
ACS Paragon Plus Environment

Page 3 of 11
ACS Medicinal Chemistry Letters
analogue 15 was found to be highly potent (IC50 = 7 nM),
CLogPb
Compound
R
X
IL-1 IC )
β (
50
1
2
3
4
5
6
7
8
equipotent to MCC950. In addition, N-cyano
sulfoximineurea derivative 16 was also found to be active
(IC50 = 21 nM). Sulfoximine moiety, the monoaza
analogues of sulfones, are stable pharmacophore which
offers a rich and versatile tool in medicinal chemistry.23
This prompted us to explore additional N-substituted
sulfoximineurea analogues. However, introduction of
alkyl, methoxy, aryl, heteroaryl, amide, ester or acyl group
at sulfoximine nitrogen proved to be detrimental to
activity (17 - 24). Overviews of all the compounds that
have been synthesized and tested for IL-1β activity are
listed in Table 1.
O
O
HO
O
1 (MCC950)
S
8 ± 0.7 nM
3.27
N
N
O
H
H
O
O
O
S
2
3
4
5
35 ± 3.2 nM
15 ± 2.1 nM
>1 µM
3.52
3.93
3.53
5.25
N
N
H
H
O
O
O
O
S
N
N
H
H
CN
N
N
H
H
CF3
O
O
9
>1 µM
N
H
N
H
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
O
6
7
>1 µM
>1 µM
4.39
2.31
N
N
H
H
O
O
Notably, 15 and 16 were highly selective against TNF-α
(IC50 >10 µM). The production of TNF-α was not affected
by 15 even at high concentration (refer supporting
information), indicating its selective effect on the NLRP3
inflammasome. Moreover, 15 blocked the recruitment of
ASC during NLRP3 activation, thus providing further
evidence to the mechanistic hypothesis of our novel N-
O
O
S
N
N
H
H
O
O
O
O
O
S
S
8
9
>1 µM
>1 µM
2.94
2.23
N
H
O
O
N
H
OH
O
O
O
cyano
sulfoximineurea
derivatives
as
NLRP3
S
10
11
>1 µM
>1 µM
4.14
4.34
N
H
inflammasome inhibitors. Furthermore, this compound
had no effect on the AIM2 inflammasome, demonstrating
specificity for NLRP3 inflammasome. The results obtained
suggested that incorporation of the N-cyano
sulfoximineurea motif may open up new directions for
further SAR development. Accordingly, we focused our
investigation on this region of the molecule, and thus, in
our optimization campaign, series of novel substituted
sulfoximineurea derivatives were synthesized and
evaluated as NLRP3 inflammasome inhibitors.
O
S
O
O
O
S
N
N
H
H
O
O
O
H
S
N
>1 µM
>1 µM
>1 µM
12
13
14
3.78
3.44
3.35
N
H
O
O
O
O
OH
P
N
H
N
H
NC
O
N
S
N
N
H
N
H
O
O
NC
S
15
16
17
18
6.8 ± 0.5 nM
2.45
N
N
H
H
O
O
N
N
O
CN
NC
Me
S
S
21.0 ± 4.0 nM
>1 µM
2.15
3.33
4.52
O
S
O
S
O
O
N
H
N
H
N
O
S
b
a
NH2
N
N
N
N
H
H
H
H
O
N
H
N
H
25
28
15
F3C
O
O
Scheme 1. Synthesis of Compound 15. Reagents and
conditions: (a) 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene
(27), n-BuLi, dry THF, -78 °C to r.t., 4 h (70%); (b)
cyanamide, MeCN, t-BuOK, NCS, r.t, 5 h (22%).
N
N
O
>1 µM
S
S
N
H
N
H
O
MeO
19
20
21
>1 µM
>1 µM
>1 µM
3.33
5.15
3.79
N
H
N
H
O
N
O
An efficient synthesis allowed rapid access to 15 as
outlined in Scheme 1. The key intermediate 25 was
obtained from commercially available p-tosyl chloride
using known procedure.24 The another key intermediate,
tricyclic amine (1,2,3,5,6,7-hexahydro-s-indacen-4-amine)
(26), and its corresponding isocyanate 27, was synthesized
via reported protocol on multi gram scale.25 The coupling
of sulfinimide 25 with 4-isocyanato-1,2,3,5,6,7-hexahydro-
s-indacene (27) provided sulfinylurea 28 in high yields,
using modified reaction conditions reported by Toth et.
al.26 Synthesis of N-substituted sulfoximines using
alkylamines27 and cyanamide28,29 are reported in
literature. However, there are no reports for the synthesis
of N-cyano sulfoximineurea albeit synthesis of
sulfinylurea followed by chlorination/amination to afford
N-alkyl sulfoximineurea are documented in literature.26
Sulfinylurea 28 was converted to N-cyano sulfoximineurea
derivative 15 using cyanamide with NCS as oxidant and
potassium tert-butoxide as base in a acetonitrile mixture.
S
F
N
H
N
H
O
N
O
S
S
N
NH
N
N
H
H
H2N
O
N
O
22
23
24
>1 µM
>1 µM
>1 µM
2.5
3.82
3.18
O
N
N
H
H
O
O
N
O
S
O
N
N
H
O
H
N
O
S
O
N
N
H
H
aIC50 values given are expressed as mean ± SEM of three
independent experiments. bCalculated from Schrödinger
Release 2018-3: QikProp, Schrödinger, LLC, New York, NY,
2018.
49
50
51
52
53
54
55
56
57
58
59
60
These results indicate that any modification of the
sulfonylurea motif in 4-14 resulted in complete loss of IL-
1β inhibition, showing the critical importance of the
sulfonylurea hydrogen bond acceptor-donor pattern.
Nevertheless, to our delight, N-cyano sulfoximineurea
ACS Paragon Plus Environment

ACS Medicinal Chemistry Letters
Page 4 of 11
Following the same reaction sequence with various bioavailability of 97%, perhaps due to good solubility. It’s
1
2
3
4
5
6
7
8
sulfinimides resulted in test compounds 16 and 29 – 34 in
good yield and high chemical purity. Synthesis of 2-24 is
described in the supporting information.
noteworthy to mention that all these compounds (15, 16
and 29-34) were highly selective against TNF-α (IC50 >10
µM). In particular, 34 show a high oral bioavailability
along with a long half-life, and represent an improvement
in PK properties with respect to regioisomers 32 and 33.
This, combined with the potency of the compound,
suggests potential for therapeutic efficacy in in vivo
models.
Table 2. Modifications on left-hand side of N-cyano-
sulfoximineurea derivatives: In vitro activity and
Pharmacokinetics propertiesa of 15, 29-34.
CN
O
9
Next, having identified compounds with high potency
and good pharmacokinetic profile, effect of the lead
compounds 15, 29 and 34, on the NLRP3 dependent
release of IL-1β was evaluated in acute in vivo LPS+ATP
challenged model in female C57BL/6 mice.31 A single oral
administration of test compounds at 10 mg/kg dose
decreases the IL-1β levels by 44%, 50% and 57%
respectively w.r.t. vehicle control (Figure 3), indicating
that even at this low dose, 15, 29 and 34 markedly reduced
the IL-1β compared to the control.
N
O
S
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
R
N
N
H
H
AUC
(µg.h/mL)b
IL-1β
IC50 (nM)
t½ (h)b
2.91
CLogPc
2.45
Compound
R
% F
56
15
29
30
6.8 ± 0.5
4.2
3.2
0.8
F
5.3 ± 0.2
2.38
59
2.61
34.0 ± 3.5
1.03
27
1.33
NC
31
11.7 ± 0.3
1.2
1.13
40
1.37
CN
32
33
34
110 ± 7
36.3 ± 3.3
23.0 ± 2.7
NC
1.28
8.2
NC
1.66
5.2
NC
24
1.18
1.19
1.48
N
N
N
97
b
aCompounds dosed 1 mg/kg iv and 3 mg/kg po. po. Mouse
PK data is mean data because of composite study design, n =
3/time point. Formulation: PO, 1% Tween 80 + 99% (0.5%)
methyl cellulose in water; IV, 5% NMP + 5% solutol + 90%
c
normal saline. Calculated from Schrödinger Release 2018-3:
QikProp, Schrödinger, LLC, New York, NY, 2018. NC: Not
calculated.
Figure 3. In vivo IL-1β inhibition of compounds 15, 29 and 34
in C57 mice. (n = 6 animals/group; female C57 mice; po 10
mg/kg; formulation, 1% Tween 80 and 99% methyl cellulose
(0.5%); ***P < 0.001 versus control, **P < 0.01 versus control;
error bar indicates SEM).
Extensive SAR work on the left-hand side aryl ring
fragment was also pursued.30 Key examples selected from
a large set of modifications are presented in Table 2.
Notably, the fluorinated compound 29 displayed excellent
potency (IC50 = 5.3 nM) as well as good pharmacokinetic
properties with AUC = 3.2 μg.h/mL at 3 mg/kg, and mice
oral bioavailability of 59%. Additional modifications on
the left-hand side aryl ring, such as addition of electron-
withdrawing groups (30 and 31), were made to potentially
improve the pharmacokinetic profile. The 4-cyano
analogue 30 displayed IL-1β inhibition IC50 of 34 nM.
However, pharmacokinetic profile of this compound was
comparatively inferior (AUC = 0.8 μg.h/mL at 3 mg/kg,
t1/2= 1 h, mice oral bioavailability 27%). The 3-cyano
analogue 31 displayed IC50 of 12 nM, with similar
pharmacokinetic profile (AUC = 1.2 μg.h/mL at 3 mg/kg,
mice oral bioavailability 40%). Next, in a quest to explore
heteroaryl analogues, the pyridyl compounds were
investigated. The 4-pyridyl analogue 32 was found to be
relatively less potent (IC50 = 112 nM). The regioisomer, 3-
pyridyl compound 33 helped to regain in vitro potency but
was accompanied by loss of plasma levels and
bioavailability. In vitro potency was further improved for
2-pyridyl compound 34 (IC50 = 23 nM). In addition, the PK
properties of 34 were also dramatically improved, with
AUC = 8.2 μg.h/mL at 3 mg/kg, t1/2= 5.2 h and mice oral
In conclusion, through systematic exploration of
rationally designed mimics of sulfonylurea moiety, we
have identified a novel series of N-cyano sulfoximineurea
derivatives as potent and selective NLRP3 inflammasome
inhibitors. The in vitro inhibition potency of N-cyano
sulfoximineurea analogue 15, on IL-1β release by LPS/ATP
stimulation was found to be similar to the reference
compound MCC950, thus indicating that structural
modifications at the sulfonylurea moiety can be tolerated.
Further 15, 29 and 34 exhibited significant IL-1β lowering
efficacy in animal models with desired pharmacokinetic
profile. These data also warrant further investigation of 15
and analogues for treatment of chronic inflammatory
diseases mediated via NLRP3 inflammasome. These NLRP3
inflammasome inhibitors with novel chemical structure
having high potency and desired PK/PD profile would be
helpful in further investigations as potential clinical
candidates.
ASSOCIATED CONTENT
Supporting information
ACS Paragon Plus Environment

Page 5 of 11
ACS Medicinal Chemistry Letters
9. Mangan, M. S. J.; Olhava, E. J.; Roush, W. R.; Seidel, H.
Supporting information file with experimental procedures
M.; Glick, G. D.; Latz, E. Targeting the NLRP3
inflammasome in inflammatory diseases. Nat Rev.
Drug Discov.2018, 17, 588–606.
1
2
3
4
5
6
7
8
and analytical data. Spectral data file. This material is
available free of charge via the Internet at
http://pubs.acs.org.
10. Dinarello, C. A.; Simon, A.; van der Meer, J. W.
Treating inflammation by blocking interleukin-1 in a
broad spectrum of diseases. Nat. Rev. Drug Discovery
2012, 11, 633−652.
11. Redondo-Castro, E.; Faust, D.; Fox, S.; Baldwin, A. G.;
Osborne, S.; Haley, M. J.; Karran, E.; Nuthall, H.;
Atkinson, P. J.; Dawson, L. A.; Routledge, C.; Allan, S.
M.; Freeman, S.; Brownlees, J.; Brough, D.
Development of a characterized tool kit for the
interrogation of NLRP3 inflammasome-dependent
responses. Sci Rep. 2018; 8, 5667.
12. Bachove, I.; Chang, C.; Anakinra and related drugs
targeting interleukin-1 in the treatment of cryopyrin-
associated periodic syndromes. Open Access
Rheumatology: Research and Reviews. 2014, 6, 15-25.
13. Zahid, A.; Li, B.; Kombe, A. J. K.; Jin, T.; Tao, J.
AUTHOR INFORMATION
Corresponding Author
*E-mail: sameeragarwal@zyduscadila.com; or
Sameer_ag@yahoo.com (SA);
Rajiv.Sharma@zyduscadila.com (RS); Fax: +91-2717-665355;
Tel: +91-2717-665555;
Conflict of interest: Authors declare no conflict of interest.
Funding Sources: No external funding.
Notes
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
ZRC Communication No. 595.
ACKNOWLEDGMENT
Authors thank Mr. Jeevan Kumar for LogP calculations using
QikProp, Schrödinger Software.
Pharmacological
Inflammasome. Front. Immunol. 2019, 10, 2538.
Inhibitors
of
the
NLRP3
14. Jiang, H.; He, H.; Chen, Y.;Huang, W.; Cheng, J.; Ye, J.;
Wang, A.; Tao, J.; Wang, C.; Liu, Q.; Jin, T.; Jiang, W.;
Deng, X.; Zhou, R. Identification of a selective and
direct NLRP3 inhibitor to treat inflammatory disorders.
J. Exp. Med. 2017, 214, 3219–3238.
15. He, Y.; Varadarajan, S.; Munoz-Planillo, R.; Burberry,
A.; Nakamura, Y.; Nunez, G. 3,4-Methylenedioxy-beta-
nitrostyrene inhibits NLRP3 inflammasome activation
by blocking assembly of the inflammasome. J. Biol.
Chem. 2014, 289, 1142–1150.
16. Coll, R. C.; Robertson, A. A.; Chae, J. J.; Higgins, S. C.;
Munoz-Planillo, R.; Inserra, M. C.; Vetter, I.; Dungan,
L. S.; Monks, B. G.; Stutz, A.; Croker, D. E.; Butler, M.
S.; Haneklaus, M.; Sutton, C. E.; Nunez, G.; Latz, E.;
Kastner, D. L.; Mills, K. H.; Masters, S. L.; Schroder, K.;
ABBREVIATIONS
NLRP3, NOD-like receptor family, pyrin domain-containing
protein 3; ASC, apoptosis-associated speck-like protein
containing a CARD; AIM2, absent in melanoma 2; DAMP,
damage-associated
molecular
pattern;
PAMP,
pathogen-associated molecular pattern; TLR, Toll-like
receptor; IFI16, Gamma-interferon-inducible protein 16; IL-
1R, Interleukin-1 receptor; LPS, Lipopolysaccharide; ATP,
adenosine triphosphate; CAPS, cryopyrin associated periodic
syndromes; TRAPS, Tumor necrosis factor receptor-
associated periodic syndrome; NCE, New Chemical Entity;
NMP, N-Methyl-2-Pyrrolidone; NCS, N-Chlorosuccinimide;
PK/PD,
Pharmacokinetic/pharmacodynamics;
TNF-α,
Cooper, M. A.; O’Neill, L. A.
A small-molecule
Tumour Necrosis Factor alpha.
inhibitor of the NLRP3 inflammasome for the
treatment of inflammatory diseases. Nat. Med. 2015, 21,
248−255.
REFERENCES
17. Baldwin, A. G.; Rivers-Auty, J.; Daniels, M. J. D.; White,
C. S.; Schwalbe, C. H.; Schilling, T.; Hammadi, H.;
Jaiyong, P.; Spencer, N. G.; England, H.; Luheshi, N.
M.; Kadirvel, M.; Lawrence, C. B.; Rothwell, N. J.;
Harte, M. K.; Bryce, R. A.; Allan, S. M.; Eder, C.;
Freeman, S.; Brough, D. Boron-based inhibitors of the
nlrp3 inflammasome. Cell Chem Biol 2017, 24, 1321–1335
e1325.
18. Guo, C.; Fulp, J. W.; Jiang, Y.; Li, X.; Chojnacki, J. E.;
Wu, J.; Wang, X. Y.; Zhang, S. Development and
characterization of a hydroxyl-sulfonamide analogue,
5-chloro-n-[2-(4-hydroxysulfamoyl-phenyl)-ethyl]-2-
methoxy-benzamide, as a novel nlrp3 inflammasome
inhibitor for potential treatment of multiple sclerosis.
ACS Chem. Neurosci. 2017, 8, 2194-2201.
19. Marchetti, C.; Swartzwelter, B.; Gamboni, F.; Neff, C.
P.; Richter, K.; Azam, T.; Carta, S.; Tengesdal, I.;
Nemkov, T.; D’Alessandro, A.; Henry, C.; Jones, G. S.;
Goodrich, S. A.; St. Laurent, J. P.; Jones, T. M.; Scribner,
C. L.; Barrow, R. B.; Altman,R. D.; Skouras, D. B.;
Gattorno, M.; Grau, V.; Janciauskiene, S.; Rubartelli, A.;
Joosten, L. A. B; Dinarello, C. A. OLT1177, a beta-
sulfonyl nitrile compound, safe in humans, inhibits the
NLRP3 inflammasome and reverses the metabolic cost
of inflammation. PNAS 2018, 115, E1530–E1539.
1.
Guo, H.; Callaway, J. B.; Ting, J. P. Inflammasomes:
mechanism of action, role in disease, and therapeutics.
Nat. Med. 2015, 21, 677−687.
2. Martinon, F.; Mayor, A.; Tschopp, J. The
inflammasomes: guardians of the body. Annu. Rev.
Immunol. 2009, 27, 229–265.
3. He, Y.; Hara, H.; Nunez, G. Mechanism and regulation
of nlrp3 inflammasome activation. Trends Biochem. Sci.
2016, 41, 1012−1021.
4. Shao, B. Z.; Xu, Z. Q.; Han, B. Z.; Su, D. F.; Liu, C. Nlrp3
inflammasome and its inhibitors: A review. Front.
Pharmacol. 2015, 6, 262-271.
5. Chen, G.; Shaw, M. H.; Kim, Y. G.; Nuñez, G. NOD-like
receptors: role in innate immunity and inflammatory
disease. Annu. Rev. Pathol. 2009, 4, 365–398.
6. Strowig, T.; Henao-Mejia, J.; Elinav, E.; Flavell, R.
Inflammasomes in health and disease. Nature 2012,
481, 278−286.
7. Zhen, Y.; Zhang, H. NLRP3 inflammasome and
inflammatory bowel disease. Front. Immunol. 2019, 10,
276.
8. Shen, H.-H.; Yang, Y.-X.; Meng, X.; Luo, X.-Y.; Li, X.M.;
Shuai, Z.-W.; Ye, D.Q.; Pan, H.F. NLRP3: A promising
therapeutic target for autoimmune diseases.
Autoimmun Rev 2018, 17, 694-702.
ACS Paragon Plus Environment

ACS Medicinal Chemistry Letters
Page 6 of 11
20. He, H.; Jiang, H.; Chen, Y.; Ye, J.; Wang, A.; Wang, C.;
Liu, Q.; Liang, G.; Deng, X.; Jiang, W.; Zhou, R.
Oridonin is a covalent nlrp3 inhibitor with strong anti-
inflammasome activity. Nat. Commun. 2018, 9, 2550.
21. Huang, Y.; Jiang, H.; Chen, Y.; Wang, X.; Yang, Y.; Tao,
J.; Deng, X.; Liang, G.; Zhang, H.; Jiang, W.; Zhou, R.
Tranilast directly targets nlrp3 to treat inflammasome-
driven diseases. EMBO Mol. Med. 2018, 10, e8689-8704.
22. Waring, M. J.; Arrowsmith, J.; Leach, A. R.; Leeson, P.
D.; Mandrell, S.; Owen, R. M.; Pairaudeau, G.; Pennie,
W. D.; Pickett, S. D.; Wang, J.; Wallace, O.; Weir, A. An
analysis of the attrition of drug candidates from four
major pharmaceutical companies. Nat. Rev. Drug
Discovery 2015, 14, 475−486.
23. Lücking, U. Sulfoximines: A neglected opportunity in
medicinal chemistry. Angew. Chem. Int. Ed. 2013, 52, 2–
12.
24. Krasnova, L. B.; Yudin, A. K. p-Tolylsulfinyl amides:ꢀ
reagents for facile electrophilic functionalization of
olefins. J. Org. Chem. 2004, 69 (7), 2584–2587.
1
2
3
4
5
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
25. Urban, F. J.; Jasys, V. J.; Raggon, J. W.; Buzon, R. A.;
Hill, P. D.; Eggler, J. F.; Weaver, J. D. Novel synthesis of
1-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)-3-[4-(1-
hydroxy-1-methyl-ethyl)-furan-2-sulfonyl]-urea,
an
anti-inflammatory agent. Synth. Commun. 2003, 33,
2029−2043.
26. Toth, J. E.; Grindey, G. B.; Ehlhardt, W. J.; Ray, J. E.;
Boder, G. B.; Bewley, J. R.; Klingerman, K. K.; Gates, S.
B.; Rinzel, S. M.; Schultz, R. M.; Weir, L. C.; Worzalla,
J. F. Sulfonimidamide analogs of oncolytic
sulfonylureas. J. Med. Chem. 1997, 40, 1018−1025.
27. Dannenberg, C. A.; Bizet, V.; Bolm, C. Direct access to
n-alkylsulfoximines from sulfides by
a sequential
imidation/oxidation procedure. Synthesis 2015, 47,
1951-1959.
28. Lafitte, G.; Parnet, V.; Pierre, R.; Raffin, C.; Vatinel, R.;
Musicki, B.; Tomas, L.; Bouix-Peter, C.; Ouvry, G.;
Daver, S.; Arlabosse, J.-M.; Boiteau, J.-G.; Harris, C. S.
Route scouting and optimization of
a
potent
sulfoximine-based inverse agonist of RORγt.
Tetrahedron 2018, 74, 5974-5986.
29. Mancheño, O. G.; Bolm, C. Synthesis of
sulfonimidamides from sulfinamides by oxidation with
N-chlorosuccinimide. Beilstein J. Org. Chem. 2007, 3,
No. 25.
30. Sharma, R.; Iyer, P.; Agarwal, S.; Novel substituted
sulfoximine compounds. PCT Int. Appl. WO
2018/225018, 13 December 2018 (Appl. PCT/IB2018/
054134, 08.06.2018).
31. Perregaux, D. G.; Mcniff, P.; Laliberte, R.; Hawryluk,
N.; Peurano, H.; Stam, E.; Eggler, J.; Griffiths, R.;
Dombroski, M. A.; Gabel, C. A. Identification and
characterization of a novel class of interleukin-1 post-
translational processing inhibitors. J. Pharmacol. Exp.
Ther. 2001, 299, 187–197.
ACS Paragon Plus Environment

Page 7 of 11
ACS Medicinal Chemistry Letters
1
2
3
4
5
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
ACS Paragon Plus Environment

ACS Medicinal Chemistry Letters
Page 8 of 11
1
2
3
4
5
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
Figure 1. Selected known NLRP3 Inflammasome Inhibitors.
133x75mm (300 x 300 DPI)
ACS Paragon Plus Environment

Page 9 of 11
ACS Medicinal Chemistry Letters
1
2
3
4
5
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
Figure 2. Schematic representation of ligand optimization.
106x31mm (300 x 300 DPI)
ACS Paragon Plus Environment

ACS Medicinal Chemistry Letters
Page 10 of 11
1
2
3
4
5
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
Scheme 1. Synthesis of Compound 15. Reagents and conditions: (a) 4-isocyanato-1,2,3,5,6,7-hexahydro-s-
indacene (27), n-BuLi, dry THF, -78 °C to r.t., 4 h (70%); (b) cyanamide, MeCN, t-BuOK, NCS, r.t, 5 h
(22%).
157x31mm (300 x 300 DPI)
ACS Paragon Plus Environment

Page 11 of 11
ACS Medicinal Chemistry Letters
1
2
3
4
5
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
Figure 3. In vivo IL-1β inhibition of compounds 15, 29 and 34 in C57 mice. (n = 6 animals/group; female
C57 mice; po 10 mg/kg; formulation, 1% Tween 80 and 99% methyl cellulose (0.5%); ***P < 0.001 versus
control, **P < 0.01 versus control; error bar indicates SEM).
130x81mm (300 x 300 DPI)
ACS Paragon Plus Environment