Discovery of Biaryl Amides as Potent, Orally Bioavailable, and CNS Penetrant RORγt Inhibitors

Article abstract of DOI:10.1021/acsmedchemlett.5b00122

A novel series of biaryl amides was identified as RORγt inhibitors through core replacement of a starting hit 1. Structure-activity relationship exploration on the biaryl moiety led to discovery of potent RORγt inhibitors with good oral bioavailability and CNS penetration. Compounds 9a and 9g demonstrated excellent in vivo efficacy in EAE mice dose dependently with once daily oral administration.

Full text of DOI:10.1021/acsmedchemlett.5b00122

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Letter
Discovery of Biaryl Amides as Potent, Orally
Bioavailable and CNS Penetrant ROR#t Inhibitors
Yonghui Wang, Wei Cai, Yaobang Cheng, Ting Yang, Qian Liu, Guifeng Zhang, Qinghua Meng,
Fangbin Han, Yafei Huang, Ling Zhou, Zhijun Xiang, Yong-Gang Zhao, Yan Xu, Ziqiang Cheng ,
Sijie Lu, Qianqian Wu, Jia-Ning Xiang, John D Elliott, Steward Leung, Feng Ren, and Xichen Lin
ACS Med. Chem. Lett., Just Accepted Manuscript • Publication Date (Web): 26 May 2015
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Discovery of Biaryl Amides as Potent, Orally Bioavailable and
CNS Penetrant RORγt Inhibitors
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Yonghui Wang,*,† Wei Cai,‡ Yaobang Cheng,‡ Ting Yang,‡ Qian Liu,‡ Guifeng Zhang,‡ Qinghua
Meng,‡ Fangbin Han,‡ Yafei Huang,† Ling Zhou,‡ Zhijun Xiang,‡ YongꢀGang Zhao,‡ Yan Xu,‡ Ziqiang
Cheng,‡ Sijie Lu,‡ Qianqian Wu,‡ JiaꢀNing Xiang,‡ John D Elliott,‡ Stewart Leung,‡ Feng Ren,‡ and
Xichen Lin*,‡
†School of Pharmacy, Fudan University, 826 Zhangheng Road, Pudong, Shanghai 201203, China
‡Research and Development, GlaxoSmithKline, No. 3 Building, 898 Halei Road, Pudong, Shanghai 201203, China
KEYWORD: RORγt inhibitor, Th17 cell differentiation, biaryl amides, EAE, multiple sclerosis
ABSTRACT: A novel series of biaryl amides was identified as RORγt inhibitors through core replacement of a starting hit 1. SAR
exploration on the biaryl moiety led to discovery of potent RORγt inhibitors with good oral bioavailability and CNS penetration.
Compounds 9a and 9g demonstrated excellent in vivo efficacy in EAE mice dose dependently with once daily oral administration.
T helper (Th) 17 cells, a lineage of CD4+ effector T cells characꢀ
terized by the production of ILꢀ17A and ILꢀ17F, are pathogenic in
human autoimmune inflammatory diseases including multiple
sclerosis (MS).[1ꢀ4] The presence of ILꢀ17 was detected in MS
lesions, and Th17 cells were observed in the infiltrations of mouse
experimental autoimmune encephalomyelitis (EAE) central nerve
system (CNS).[5,6] Differentiation and function of Th17 cells are
controlled by the transcription factor retinoic acid receptorꢀrelated
orphan receptorꢀgammaꢀt (RORγt).[7ꢀ9,11] It has been shown that
the genetic deficiency of RORγt in mice severely impaired Th17
cell differentiation and conferred resistance to EAE.[10] RORγt
inhibitors has potential utility in reducing the activity of Th17
cells and therefore can be developed as therapeutic agents for the
treatment of Th17 cell mediated autoimmune diseases.[12ꢀ18]
best RORγt potency in the FRET assay.[32,34] The heteroatom such
as oxygen and nitrogen on the ring decreased (4a) and even abolꢀ
ished (4d, 4f, 4h) RORγt activities. For the sixꢀmembered ring
analogs, paraꢀsubstituted aryl amide (4i) showed better RORγt
potency than the metaꢀsubstituted one (4g). Because of its reasonꢀ
able RORγt potency, good CNS penetration (Br/Bl = 2.0), and
improved ligand efficiency (LE) and lipophilic ligand efficiency
(LLE) (0.33 and 2.3 for 4i compared to 0.29 and 1.9 for 1, respecꢀ
tively)35 and easy modification/diversification, the aryl amide 4i
was used as the new chemistry starting point for optimization.
A few small molecule RORγt inhibitors have been reported in
literature.[19] Digoxin,[20] SR1001[21] and Ursolic acid[22] were first
reported to inhibit RORγt and ameliorate EAE in mice via intraꢀ
peritoneal administration. Other small molecular RORγt inhibiꢀ
tors[23ꢀ31] were later disclosed. Recently, we reported discovery of
thiazole ketone amides (e.g., 2) and thiophene ketone amides (e.g.,
3) as novel RORγt inhibitors based on a high throughput screenꢀ
ing (HTS) hit 1 (Figure 1).[32] These ketones, especially the thioꢀ
phene ketones, showed good RORγt activities but were poorly
orally bioavailable and lack of CNS penetration that is believed to
be important for developing an effective oral MS drug. In this
paper, we report the discovery of novel biaryl amides as first poꢀ
tent, orally bioavailable and CNS penetrant RORγt inhibitors
which demonstrated EAE in vivo efficacy dose dependently via
oral administration.
Figure 1. Structures of RORγt inhibitors (1ꢀ3)
In order to explore SAR of the biaryl moiety of the amide, a verꢀ
satile synthesis of the general structures of biaryl amides was
developed (Scheme 1).[36] Biaryl amines 7 were prepared from
either bromoanilines 5 through Suzuki coupling with aryl boronic
acids, or from reaction of aryl bromides with aniline boronic esꢀ
ters 6, obtained from 5. Coupling 7 with acids A, or acid chlorides
B, or perfluorophenyl esters C afforded the desired biaryl amides
8 or 9. The biaryl amides could also be prepared by first coupling
of 5 with A to form amides 10, which were converted to the target
compounds directly via Suzuki coupling, or via its boronic ester
intermediate 11.
The lack of CNS penetration of thiazole/thiophene ketones was
attributed to their ketone moiety as the nonꢀketone thiazole amide
1 is CNS penetrant with a brainꢀtoꢀblood ratio (Br/Bl) of 1.5 in a
mouse CNS study (i.p., 2mg/kg).[33] Encouraged by the CNS data
of 1, we conducted thiazole core replacement with a number of
different aromatic rings (4aꢀ4i), aiming to identify a suitable scafꢀ
fold for multiꢀproperty optimization (Table 1). Among the fiveꢀ
membered ring analogs, 2,4ꢀsubstituted thiophene 4c showed the
We investigated the binding mode of compound 4i and its derivaꢀ
tives in RORγt LBD based on the coꢀcrystal structure of a similar
aryl amide with RORγt LBD (pdb code: 4NIE).[37] The perpendicꢀ
ular confirmation of the two aryl rings in the left hand side (LHS)
of the amides provided preferred interꢀmolecular interactions with
the surrounding hydrophobic residues in the RORγt LBD and was
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believed to be important for the RORγt binding affinity (Figure 1).
significantly decreased both RORγt and Th17 potency. Although
1
2
3
4
5
6
7
8
Subsequently, the substitutions on the orthoꢀpositions of the two
aryl rings, which force the two aryls to take perpendicular conꢀ
formation, were studied extensively and the key SAR of the biarꢀ
yls was summarized in Table 2. Nonꢀsubstituted biphenyl amide
8a showed a RORγ FRET pIC50 of 6.3. Adding a Cl group on
orthoꢀposition of the central phenyl ring (8b) enhanced RORγt
activity. Keeping the orthoꢀCl on the central phenyl ring, adding a
hydrophobic group on orthoꢀposition of the terminal phenyl ring
provided potent RORγt inhibitors (8cꢀ8f) with pIC50s ≥ 8.0 in the
FRET assay. Biaryl amides 8cꢀ8f also showed good cellular activꢀ
ities in the Th17 cell differentiation assay (pIC50 > 6.0).[32ꢀ34] Obꢀ
viously, the cLogP of 8cꢀ8f are relatively high (4.4~5.1, from
ChemBioDraw Ultra 12.0). Replacing ꢀOiPr moiety (8f) with ꢀ
CH2NMe2 (8g, cLogP 3.7) or changing the phenyl ring to a pyriꢀ
dine ring (8h, cLogP 4.0) or other heteroꢀaromatic rings such as
pyrole (8i, cLogP 3.5) lowered RORγt potency, which also resultꢀ
ed in essentially no activity in Th17 cell differentiation assay.
These findings indicate that the binding pocket where the LHS
aryl occupies is hydrophobic and unable to tolerate some polar
moieties.
polar groups like carboxylic acid (8o) decreased the RORγt poꢀ
tency dramatically, certain polar groups such as acetyl (8n) were
found to be tolerated on the central phenyl. Encouraged by this, a
number of heteroꢀaromatic rings were introduced on the central
phenyl and the resulting compounds (e.g., 8pꢀ8q) showed good
RORγt activity in both FRET and Th17 assays. It is good to see
that the cLogP of 8p and 8q are relatively lower (3.6 and 3.0,
respectively), resulting in higher LLE values (4.7 and 5.4, respecꢀ
tively) although their molecular weights are higher.
9
Scheme 1. General synthetic procedures for biaryl amidesa
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X
O
O
ArBr
b
B
NH2
X
Y
Y
6
Ar
NH2
a
A or B
or C
c
X
7
ArB(OH)2
b
Br
NH2
O
R1
O
X
S
5
Y
Ar
O
d
A
Y
N
H
O
O
X
ArB(OH)2
f
Table 1. SAR of thiazole core replacements
S
8 or 9
Br
R1
O
Y
N
H
10
e
O
R1
O
S
O
R1
O
X
O
ArBr
f
S
B
O
R2
A (R2 = OH), B (R2 = Cl), C (R2
F
O
O
a
O
F
F
F
RORγ FRET pIC50
Y
N
H
Compd
R
=
)
(% max inhibitionb)
6.0±0.08 (116)
5.1±0.06 (80)
6.4±0.06 (105)
7.1±0.01 (116)
< 4.6
F
11
aReagents and conditions: (a) Bis(pinacolato)diborane, PdCl2(dppf), KOAc,
DMF, 100 ° C. (b) triꢀtertꢀbutyl phosphine (tetrafluoroboric acid salt),
Pd2(dba)3, Na2CO3, dioxane, 100 °C, microwave. (c) For acid A, EDC, HOBt,
DCM; For acid chloride B, triethylamine, DCM; For perfluorophenyl ester C,
DIPEA, DCM, RT. (d) EDC, HOBt, DCM; or HATU, DIPEA, DCM. (e)
1
4a
4b
4c
4d
4e
4f
Bis(pinacolato)diborane, PdCl2(dppf), KOAc, DMF, 100
° C; or
Bis(pinacolato)diborane, Pd2(dba)3, tricyclohexylphosphine, KOAc, dioxane,
90 °C. (f) PdCl2(dppf), Cs2CO3, CH3CN, water, 100 °C, microwave; or
Pd(PPh3)4, Na2CO3, dioxane, water, 100 °C, microwave.
Table 2. Biaryl SAR of the amides
6.3±0.04 (120)
< 4.6
O
O
S
S
O
Ar1
Ar2
N
H
6.2±0.02 (120)
4g
RORγ FRET
Th17
pIC50
a
Compd
Ar1
Ar2
pIC50 (% max
a
inhibitionb)
< 4.6
4h
4i
6.3±0.30 (90)
<5
8a
8b
7.0±0.06 (107)
7.8±0.04 (109) 5.2
8.3±0.02 (100) 6.0
8.2±0.02 (106) 6.7
apIC50 value is the average of at least two determinations, the error
expressed by ±SEM; b% max inhibition measured against activation
by the surrogate agonist.
8c
8d
8e
We then fixed the OCF3 group at orthoꢀposition of the LHS terꢀ
minal aryl and studied SAR of substitution on the central phenyl
ring of the amides (Table 2). Adding a hydrophobic group such as
methyl (8l) or CF3 (8m) on the central phenyl ring increased Th17
potency while replacing the central phenyl ring with pyridine (8k)
8.5±0.10 (98)
7.1
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introduction of two more Hꢀbond donors as well as increase of
8.0±0.00 (120) 7.1
6.0±0.00 (117) <5
8f
1
2
3
4
5
6
7
8
topological polar surface area (tPSA) in 9b. Switching OCF3 (9a)
to OCF2 (9c) lowered its CNS penetration. The CNS penetration
was further decreased when OCF2 (9c) was replaced by a CN
group (9d). With a Cl group in the paraꢀposition of LHS phenyl
and only one substituent (F, Me, or Cl) in the ortho position of
central phenyl, compounds (9fꢀ9h) showed good RORγt potency
and CNS penetration. Compared to methyl sulfone 9h, the ethyl
sulfone 9i demonstrated the best CNS penetration (Br/Bl = 2.0).
Clearly, the data of CNS penetration were well correlated to valꢀ
ues of tPSA and/or cLogP. As a result, LLE value is relatively
low for those biaryl amides with better CNS penetration (Table 3).
8g
7.2±0.00 (106) <5.3
6.8±0.07 (115) <5.2
8.1±0.00 (113) 6.8
6.4±0.12 (106) 5.1
8.3±0.04 (112) 7.6
8.2±0.02 (113) 6.9
8h
8i
9
8j
10
11
12
13
14
15
16
17
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N
8k
8l
8m
8.0±0.08 (101) 5.6
8n
8o
5.1±0.05 (68)
8.3±0.11 (96)
<5
Figure 1 Predicted binding mode of compound 4i (brown) and
structural overlay with a previously published tertiary amine (blue)
coꢀcrystal structure with RORγt LBD using SurflexꢀDock v2.3 in
Sybyl 8.1.[37]
7.2
8p
Table 4. Mouse PKa of the RORγt inhibitors
8.4±0.01 (112) >7.5
8q
8r
iv, 1 mg/kgb
po, 2 mg/kgc
Comp
d
T1/
8.2±0.08 (96)
8.4±0.12 (93)
8.4±0.02 (97)
7.5
Clb
Vss
Cmax
DNAUC0~∞
F
2
(mL/min/kg (L/kg (ng/mL (ng.h/mL)/(mg/kg (%
(h)
)
)
)
)
)
>8.5
>8.2
8s
8t
8d 2.2
8e 4.2
9a 9.7
9g
17.6
3.2 210.7
3.8 202.7
4.4 213.5
713
75
10
2
10
0
11.6
5.5
1313
2465
4048d
apIC50 value is the average of at least two determinations, the error
expressed by ±SEM (for FRET assay) ; b% max inhibition measured
against activation by the surrogate agonist.
b
aMale C57BL/6 mice; iv formulation: DMSO: 10% hydroxypropylꢀ
c
βꢀcyclodextrin = 1:99 (w:v); po formulation: DMSO: 1% methylꢀ
cellulose (W/V) =1:99; for 9a, DMSO: 10% hydroxypropylꢀβꢀ
cyclodextrin; d10 mg/kg (po).
We next added a second substituent to the central phenyl ring to
constrain the preferred perpendicular conformation. As expected,
additional substituent CN (8r), Me (8s) or Cl (8t) boosted RORγt
activities in both FRET and Th17 assays.
Several representative compounds were evaluated for their mouse
PK profile (Table 4). Biaryl amides 8d, 8e and 9a demonstrated
good PK profile with oral bioavailabilities of 75%, 102% and
100%, respectively. Compound 9g was only evaluated via po
administration and showed excellent oral exposure.
Compound 8t was used as a tool compound for RORγt biological
studies because of its excellent in vitro activities as well as good
oral exposure and CNS penetration.[38] Encouraged by the profile
of 8t, we incorporated the previous SAR learnings and further
optimized the LHS biaryl part as well as right hand side (RHS)
sulfone part of the amides, trying to obtain a molecule with more
balanced profile (Table 3). Changing the ethyl sulfone in 8t with a
methyl sulfone (9a) resulted in a similar RORγt potency and CNS
penetration. However, replacing the methyl sulfone with a primaꢀ
ry sulfonamide (9b) basically eliminate the CNS penetration altꢀ
hough the RORγt and Th17 potency remained, possibly due to
With good Th17 activity and mouse oral exposure, we then evaluꢀ
ated 9a and 9g in EAE mice where Th17 cells play a critical role
(Figure 2).[33] Compounds 9a and 9g were orally administered
once daily at 3 doses (1, 3, 10 mg/kg) to EAE mice from the day
of immunization. Compared to the control, the treatment with 9a
or 9g resulted in a delay and significant reduction in clinical seꢀ
verity of EAE in a dose dependent manner. Compared to thiazole
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Table 3. SAR of the biaryl amides
1
2
3
4
O
R2
O
S
O
5
6
N
H
7
8
9
RORγ FRET
Th17
pIC50
Br/Blc
(AUCbrain/AUCblood
pIC50a (% max
inhibitionb)
Compd
R1
Z
X
Y
R2
tPSAd cLogPd LLEe
a
)
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
8t
H
H
OCF3
OCF3
Cl
Cl
Cl
Cl
Et
8.4±0.02 (97)
8.3±0.15 (96)
>8.2
0.78 (946/1220)
72.5
72.5
5.2
4.7
3.2
3.6
9a
Me
7.4
0.79 (658/835)
9b
9c
9d
9e
9f
H
H
OCF3
OCF2
CN
Cl
Cl
Cl
Cl
F
Cl
Cl
Cl
Cl
H
NH2 8.5±0.11 (108)
8.1
8.0
7.1
6.8
7.3
7.2
7.6
8.1
0.08 (388/4878)
0.39 (462/1182)
0.10 (200/1928)
0.06 (76/1378)
98.5
72.5
87.0
87.0
72.5
72.5
72.5
72.5
4.5
4.6
4.2
4.3
4.7
4.5
5.0
5.5
4.0
3.9
4.0
4.1
3.3
3.7
3.1
2.7
Et
Et
8.5±0.26 (107)
8.2±0.11 (94)
8.4±0.03 (96)
8.0±0.28 (101)
8.2±0.08 (92)
8.1±0.09 (98)
8.2±0.01 (99)
H
F
CN
Et
Cl
Cl
Cl
Cl
OCF3
OCF3
OCF3
OCF3
Me
Me
Me
Et
1.17 (2354/2017)
1.47 (3517/2397)
0.98 (1696/1729)
2.0 (1764/881)
9g
9h
9i
Me
Cl
Cl
H
H
H
apIC50 value is the average of at least two determinations, the error expressed by ±SEM (for FRET assay) ; b% max inhibition measured
against activation by the surrogate agonist; cbrain to blood ratio33; dobtained from ChemBioDraw Ultra 12.0; eLLE = pIC50ꢀcLogP.35
ketone amide 2 which only showed EAE efficacy up to day 20 at
100mg/kg twice daily dosing,[32] the biaryl amides 9a and 9g are
much more efficacious. This could be attributed to their good in
vitro activities as well as much improved oral exposure and CNS
penetration. However, it should be noted that although 9g had
more brain exposure than 9a, it exhibited less efficacy than 9a in
EAE experiments, indicating that there might be additional factors
such as “free” brain concentration affecting in vivo efficacy.
Figure 3. (a) Treatment efficacy of compound 9a in mouse EAE
in different doses (1, 3, 10mg/kg, p.o., q.d.). (c) Treatment efficaꢀ
cy of compound 9g in mouse EAE in different doses (1, 3,
10mg/kg, p.o., q.d.). Repeated ANOVA, followed by Dunnett's
Multiple
Comparison
Test
was
apꢀ
plied,*p<0.05,**p<0.01,***p<0.001. Notes:
In summary, we have discovered a novel series of biaryl amides
as RORγt inhibitors. Detailed SAR study on the LHS biaryl moieꢀ
ty of the amides led to discovery of potent RORγt inhibitors with
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(9) Yang, X. O.; Pappu, B. P.; Nurieva, R.; Akimzhanov, A.; Kang,
excellent oral bioavailability and CNS penetration. The key comꢀ
H. S.; Chung, Y.; Ma, L.; Shah, B.; Panopoulos, A. D.; Schluns, K. S.;
Watowich, S. S.; Tian, Q.; Jetten, A. M.; Dong, C. T helper 17 lineage
differentiation is programmed by orphan nuclear receptors ROR alpha
and ROR gamma. Immunity. 2008, 28, 29ꢀ39.
(10) Wang, X.; Zhang, Y.; Yang, X. O.; Nurieva, R. I.; Chang, S.
H.; Ojeda, S. S.; Kang, H. S.; Schluns, K. S.; Gui, J.; Jetten, A. M.;
Dong, C. Transcription of Il17 and Il17f is controlled by conserved
noncoding sequence 2. Immunity. 2012, 36, 23ꢀ31.
(11) Wang, Y.; Kumar, N.; Solt, L. A.; Richardson, T. I.; Helvering,
L. M.; Crumbley, C.; GarciaꢀOrdonez, R. D.; Stayrook, K. R.; Zhang,
X.; Novick, S.; Chalmers, M. J.; Griffin, P. R.; Burris, T. P. Modulaꢀ
tion of retinoic acid receptorꢀrelated orphan receptor alpha and gamꢀ
ma activity by 7ꢀoxygenated sterol ligands. J. Biol. Chem. 2010, 285,
5013ꢀ5025.
(12) Huh, J. R.; Littman, D. R. Small molecule inhibitors of RORγt:
targeting Th17 cells and other applications. Eur. J. Immunol. 2012, 42,
2232ꢀ2237.
(13) Burris, T. P.; Busby, S. A.; Griffin, P. R. Targeting orphan nuꢀ
clear receptors for treatment of metabolic diseases and autoimmunity.
Chemistry & Biology. 2012, 19, 51ꢀ59.
(14) Murali Dhar, T. G.; Zhao, Q.; Markby, D. W. Targeting the
nuclear hormone receptor RORγt for the treatment of autoimmune
and inflammatory disorders. Annu. Rep. Med. Chem., 2013, 48, 169ꢀ
182.
(15) Lee, J. S.; Cua, D. J. Emerging Landscape of RORγt Biology.
Immunity, 2014, 40, 451ꢀ452.
(16) Yang, J.; Sundrud, M. S.; Skepner, J.; Yamagata, T. Targeting
Th17 cells in autoimmune diseases. Trends in Pharmacological Sci-
ences, 2014, 35, 493ꢀ500.
(17) Isono, F.; FujitaꢀSata, S.; Ito, S. Inhibiting RORγt/Th17 axis
for autoimmune disorders. Drug Discovery Today, 2014, 19, 1205ꢀ
1211.
(18) Dong, C. Targeting Th17 cells in immune diseases. Cell Re-
search, 2014, 24, 901ꢀ903.
(19) Fauber, B. P.; Magnuson, S. Modulators of the nuclear recepꢀ
tor retinoic acid receptorꢀrelated orphan receptorꢀγ (RORγ or RORc).
J. Med. Chem., 2014, 57, 5871ꢀ5892.
(20) Huh, J. R.; Leung, M. W. L.; Huang, P.; Ryan, D. A.; Krout,
M. R.; Malapaka, R. R. V.; Chow, J.; Manel, N.; Ciofani, M.; Kim, S.
V.; Cuesta, A.; Santori, F. R.; Lafaille, J. J.; Xu, H. E.; Gin, D. Y.;
Rastinejad, F.; Littman, D. R. Digoxin and its derivatives suppress
TH17 cell differentiation by antagonizing RORγt activity. Nature.
2011, 472, 486ꢀ490.
(21) Solt, L. A.; Kumar, N.; Nuhant, P.; Wang, Y.; Lauer, J. L.; Liu,
J.; Istrate, M. A.; Kamenecka, T. M.; Roush, W. R.; Vidovic, D.;
Schürer, S.C.; Xu, J.; Wagoner, G.; Drew, P. D.; Griffin, P. R.; Burris,
T. P. Suppression of Th17 Differentiation and autoimmunity by a
synthetic ROR ligand. Nature. 2011, 472, 491ꢀ494.
(22) Xu, T.; Wang, X.; Zhong, B.; Nurieva, R. I.; Ding, S.; Dong,
C. Ursolic acid suppresses interleukinꢀ17 (ILꢀ17) production by selecꢀ
tively antagonizing the function of RORγt protein. J. Biol. Chem.
2011, 286, 22707ꢀ22710.
(23) Solt, L. A.; Kumar, N.; He, Y.; Kamenecka, T. M.; Griffin, P.
R.; Burris, T. P. Identification of a selective RORγ ligand that supꢀ
presses Th17 cells and stimulates T regulatory cells. ACS Chem. Biol.
2012, 7, 1515ꢀ1519.
(24) Kumar, N.; Lyda, B.; Chang, M. R.; Lauer, J. L.; Solt, L. A.;
Burris, T. P.; Kamenecka, T. M.; Griffin, P. R. Identification of
SR2211: a potent synthetic RORγꢀselective modulator. ACS Chem.
Biol. 2012, 7, 672ꢀ677.
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pounds 9a and 9g demonstrated a dose dependent EAE efficacy in
mice when administrated orally once a day. Further optimization
on sulfone moiety of the biaryl amides to balance potency and
some developability properties such as solubility is onꢀgoing.
ASSOCIATED CONTENT
Supporting Information
Synthetic procedures and compound characterization; molecular
modeling studies; mouse CNS measurement description. This
material is available free of charge via the Internet at http://
pubs.acs.org.
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AUTHOR INFORMATION
Corresponding Author
*(Y. Wang) Phone: +8621ꢀ51980118. Eꢀmail: yonghuiꢀ
wang@fudan.edu.cn; *(X. Lin) Phone: +8621ꢀ61590718. Eꢀmail:
xichen.2.lin@gsk.com
Author Contributions
The manuscript was written through contributions of all authors.
All authors have given approval to the final version of the manuꢀ
script.
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENT
We thank Wei Zhang, Jing Zhang, Gang An, Jinqiang Zhang,
Melanie Huxdorf, Yingli Ma, Shuai Wang, Hui Lei, Hongtao Lu
and Zhong Zhong for their help and discussions.
REFERENCES
(1) Dardalhon, V.; Korn, T.; Kuchroo, V. K.; Anderson, A. C. Role
of Th1 and Th17 cells in organꢀspecific autoimmunity. J. Autoimmun.
2008, 31, 252ꢀ256.
(2) Komiyama, Y.; Nakae, S.; Matsuki, T.; Nambu, A.; Ishigame,
H.; Kakuta, S.; Sudo, K.; Iwakura, Y. ILꢀ17 plays an important role in
the development of experimental autoimmune encephalomyelitis. J.
Immunol. 2006; 177, 566–573.
(3) Tzartos, J. S.; Friese, M. A.; Craner, M. J.; Palace, J.; Newꢀ
combe, J.; Esiri, M. M.; Fugger, L. Interleukinꢀ17 production in cenꢀ
tral nervous systemꢀinfiltrating T cells and glial cells is associated
with active disease in multiple sclerosis. Am. J. Pathol. 2008, 172,
146ꢀ155.
(4) Ivanov, I. I.; McKenzie, B. S.; Zhou, L.; Tadokoro, C. E.;
Lepelley, A.; Lafaille, J. J.; Cua, D. J.; Littman, D. R. The orphan
nuclear receptor RORgammat directs the differentiation program of
proinflammatory ILꢀ17+ T helper cells. Cell. 2006, 126, 1121ꢀ1133.
(5) Matusevicius, D.; Kivisäkk, P.; He, B.; Kostulas, N.; Ozenci,
V.; Fredrikson, S.; Link, H. Interleukinꢀ17 mRNA expression in
blood and CSF mononuclear cells is augmented in multiple sclerosis.
Mult. Scler. 1999, 5, 101ꢀ104.
(6) Lock, C.; Hermans, G.; Pedotti, R.; Brendolan, A.; Schadt, E.;
Garren, H.; LangerꢀGould, A.; Strober, S.; Cannella, B.; Allard, J.;
Klonowski, P.; Austin, A.; Lad, N.; Kaminski, N.; Galli, S. J.; Oksenꢀ
berg, J. R.; Raine, C. S.; Heller, R.; Steinman, L. Geneꢀmicroarray
analysis of multiple sclerosis lesions yields new targets validated in
autoimmune encephalomyelitis. Nat. Med. 2002, 8, 500ꢀ508.
(7) Solt, L. A.; Burris, T. P. Action of RORs and their ligands in
(patho)physiology. Trends Endocrinol. Metab. 2012, 23, 619ꢀ627.
(8) Manel, N.; Unutmaz, D.; Littman, D. R. The differentiation of
human T(H)ꢀ17 cells requires transforming growth factorꢀbeta and
induction of the nuclear receptor RORgammat. Nat. Immunol. 2008, 9,
641ꢀ649.
(25) Huh, J. R.; Englund, E. E.; Wang, H.; Huang, R.; Huang, P.;
Rastinejad, F.; Inglese, J.; Austin, C. P.; Johnson, R. L.; Huang, W.;
Littman, D. R. Identification of potent and selective diphenylproꢀ
panamide RORγ inhibitors. ACS Med. Chem. Lett. 2013, 4, 79ꢀ84.
(26) Fauber, B. P.; de Leon Boenig, G.; Burton, B.; Eidenschenk,
C.; Everett, C.; Gobbi, A.; Hymowitz, S. G.; Johnson, A. R.; Liimatta,
M.; Lockey, P.; Norman, M.; Ouyang, W.; Rene, O.; Wong, H. Strucꢀ
tureꢀbased design of substituted hexafluoroisopropanolꢀarylꢀ
ACS Paragon Plus Environment

ACS Medicinal Chemistry Letters
Page 6 of 7
sulfonamides as modulators of RORc. Bioorg. Med. Chem. Lett. 2013,
S.; and Lin, X. Discovery of novel Nꢀ(5ꢀ(arylcarbonyl)thiazolꢀ2ꢀ
23, 6604ꢀ6609.
yl)amides and Nꢀ(5ꢀ(arylcarbonyl)thiophenꢀ2ꢀyl)amides as potent
RORγt inhibitors. Bioorg. Med. Chem. 2014, 22, 692ꢀ702.
(33) See supporting information for the details.
1
2
3
4
5
6
7
8
(27) Zhang, Y.; Xue, X.; Jin, X.; Song, Yu; Li, J.; Luo, X.; Song,
M.; Yan, W.; Song, H.; Xu, Y. Discovery of 2ꢀoxoꢀ1,2ꢀ
dihydrobenzo[cd]indoleꢀ6ꢀsulfonamide derivatives as new RORγ
inhibitors using virtual screening, synthesis and biological evaluation.
Eur J. Med. Chem. 2014, 78, 431ꢀ441.
(28) Toyama, H.; Nakamura, Masaharu.; Nakamura, Masahiko.;
Matsumoto, Y.; Nakagomi, M.; Hashimoto, Y. Development of novel
siliconꢀcontaining inverse agonists of retinoic acid receptorꢀrelated
orphan receptors. Bioorg. Med. Chem. 2014, 22, 1948ꢀ1959.
(29) Fauber, B. P.; Rene, O.; Burton, B.; Everett, C.; Gobbi, A.;
Hawkins, J.; Johnson, A. R.; Liimatta, M.; Lockey, P.; Norman, M.;
Wong, H. Identification of tertiary sulfonamides as RORc inverse
agonists. Bioorg. Med. Chem. Lett. 2014, 24, 2182ꢀ2187.
(30) Nishiyama, Y.; Nakamura, M.; Misawa, T.; Nakagomi, M.;
Makishima, M.; Ishikawa, M.; Hashimoto, Y. Structureꢀactivity relaꢀ
tionshipꢀguided development of retinoic acid receptorꢀrelated orphan
receptor gamma (RORγ)ꢀselective inverse agonists with a phenanꢀ
thridinꢀ6(5H)ꢀone skeleton from a liver X receptor ligand. Bioorg.
Med. Chem. 2014, 22, 2799ꢀ2808.
(31) Fauber, B. P.; Rene, O.; de Leon Boenig, G.; Burton, B.; Deng,
Y.; Eidenschenk, C.; Everett, C.; Gobbi, A.; Hymowitz, S. G.; Johnꢀ
son, A. R.; La, H.; Liimatta, M.; Lockey, P.; Norman, M.; Ouyang,
W.; Wang, W.; Wong, H. Reduction in lipophilicity improved the
solubility, plasmaꢀprotein binding, and permeability of tertiary sulꢀ
fonamide RORc inverse agonists. Bioorg. Med. Chem. Lett. 2014, 24,
3891ꢀ3897.
(34) Zhang, W.; Zhang, J.; Fang, L.; Zhou, L.; Wang, S.; Xiang, Z.;
Li, Y.; Wisely, B.; Zhang, G.; An, G.; Wang, Y.; Leung, S.; Zhong, Z.
Increasing human Th17 differentiation through activation of orphan
nuclear receptor retinoid acidꢀrelated orphan receptor γ (RORγ) by a
class of aryl amide compounds. Mol. Pharmacol. 2012, 82, 583ꢀ590.
(35) Hopkins1, A. L.; Keserü, G. M.; Leeson, P. D.; Rees, D. C.;
Reynolds, C. H. The role of ligand efficiency metrics in drug discovꢀ
ery. Nature Rev. Drug Discov., 2014, 13, 105ꢀ121.
(36) Wang, Y.; Cai, W.; Liu, Q.; Meng, Q.; Cheng, Y.; Yang, T.;
Zhang, G.; Xiang, J.; Wu, C. Preparation of Nꢀsubstituted 2ꢀ[4ꢀ
(ethylsulfonyl)phenyl]acetamides as retinoidꢀrelated orphan receptor
gamma modulators. WO2013029338A1, 2013.
(37) Yang, T.; Liu, Q.; Cheng, Y.; Cai, W.; Ma, Y.; Yang, L.; Wu,
Q.; OrbandꢀMiller, L. A.; Zhou, L.; Xiang, Z.; Huxdorf, M.; Zhang,
W.; Zhang, J.; Xiang, J.; Leung, S.; Qiu, Y.; Zhong, Z.; Elliott, J. D.;
Lin, X. Discovery of tertiary amine and indole derivatives as potent
RORγt inverse agonists. ACS Med. Chem. Lett. 2014, 5, 65ꢀ68.
(38) Xiao, S.; Yosef, N.; Yang, J.; Wang, Y.; Zhou, L.; Zhu, C.;
Wu, C.; Baloglu, E.; Schmidt, D.; Ramesh, R.; Lobera, M.; Sun, M. S.;
Tsai, P.ꢀY.; Xiang, Z.; Wang, J.; Xu, Y.; Lin, X..; Kretschmer, K.;
Rahi, P. B.; Young, R. A.; Zhong, Z.; Hafler, D. A.; Regev, A.;
Ghosh, S.; Marson, A.; Kuchroo, V. K. Smallꢀmolecule RORγt antagꢀ
onists inhibit T helper 17 cell transcriptional network by divergent
mechanisms. Immunity, 2014, 40, 477ꢀ489.
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(32) Wang, Y.; Cai, W.; Zhang, G.; Yang, T.; Liu, Q.; Cheng, Y.;
Zhou, L.; Ma, Y.; Cheng, Z.; Lu, S.; Zhao, Y.ꢀG.; Zhang, W.; Xiang,
Z.; Wang, S.; Yang, L.; Wu, Q.; OrbandꢀMiller, L. A. Xu, Y.; Zhang,
J.; Gao, R.; Huxdorf, M.; Xiang, J.ꢀX.; Zhong, Z.; Elliott, J. D.; Leung,
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Control
Mouse EAE
9a, 1mg/kg, po, qd
9a, 3mg/kg, po, qd
9a, 10mg/kg, po, qd
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Treatment
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