Discovery and Assessment of Atropisomers of (±)-Lesinurad

Article abstract of DOI:10.1021/acsmedchemlett.6b00465

(+)- and (?)-Lesinurad were isolated as atropisomers from racemic lesinurad for the first time. No interconversion was observed between the two atropisomers under various conditions tested. The two atropisomers showed significant differences in hURAT1 highly expressed HEK293 cell-based inhibition assays, monkey pharmacokinetic studies, and in vitro human recombinant CYP2C9 stability studies. It was speculated that (+)-lesinurad might offer a better hyperuricemia/gout therapy than (?)-lesinurad or the racemate.

Full text of DOI:10.1021/acsmedchemlett.6b00465

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Letter
Discovery and Assessment of Atropisomers of (±)-Lesinurad
Jianfei Wang, Wenqin Zeng, Shaohua Li, Liang Shen, Zhengxian
Gu, Yang Zhang, Jian Li, Shuhui Chen, and Xiangbo Jia
ACS Med. Chem. Lett., Just Accepted Manuscript • DOI: 10.1021/acsmedchemlett.6b00465 • Publication Date (Web): 14 Feb 2017
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Discovery and Assessment of Atropisomers of (±)-Lesinurad
Jianfei Wang,† Wenqin Zeng,† Shaohua Li,† Liang Shen,† Zhengxian Gu,† Yang Zhang,*,† Jian Li,†
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Shuhui Chen,† and Xiangbo Jia‡
†WuXi AppTec, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, PR China
‡
Sagacity New Drug R&D Co., Ltd., 18 Zhenze Road, Xinwu District, Wuxi, Jiangsu, 214135, PR China
KEYWORDS: Atropisomer, Lesinurad, Gout, Hyperuricemia
ABSTRACT: (+)ꢀ and (ꢀ)ꢀlesinurad were isolated as atropisomers from racemic lesinurad for the first time. No interconversion
was observed between the two atropisomers under various conditions tested. The two atropisomers showed significant differences
in hURAT1 highly expressed HEK293 cellꢀbased inhibition assays, monkey pharmacokinetic studies and in vitro human recombiꢀ
nant CYP2C9 stability studies. It was speculated that (+)ꢀlesinurad might offer a better hyperuricemia/gout therapy than (ꢀ)ꢀ
lesinurad or the racemate.
Gout is a crystalꢀdeposition arthritis caused by super saturaꢀ
tion and precipitation of monosodium urate (MSU) in tissues
or joints, which affects over 20 million people worldwide.1ꢀ5
Hyperuricemia is the biological fundamental of gout.6 It is a
purine related metabolic disease defined as serum Uric Acid
(sUA) level >6.8 mg/dL.7 The standard medical treatment of
hyperuricemia is urateꢀlowing therapy (ULT) with a goal to
lower sUA level below 6 mg/dL.4,5,8 Several ULTs are availaꢀ
ble, including xanthineꢀoxidase inhibitors (XOIs, e.g., allopuꢀ
rinol and febuxostat) to reduce sUA formation, and uric acid
reꢀabsorption inhibitors (URIs, e.g., benzbromarone and proꢀ
benecid) to increase the excretion of uric acid. In recent years,
selectively blocking hURAT1 (human uric acid transporter 1,
SLC22A12) to reduce reꢀabsorption of uric acid at kidney tuꢀ
bules gained attention for the treatment of hyperuricemia and
gout.9ꢀ12 Among them, Zurampic (lesinurad, Scheme 1), a firstꢀ
inꢀclass selective hURAT1 inhibitor, was approved by the
FDA as an oral therapy for hyperuricemia and gout in late
2015. The approved and recommended dose of Zurampic is
200 mg once daily in combination with allopurinol or febuxoꢀ
stat.13,14 The 400 mg dose of lesinurad, albeit demonstrating
good therapeutic efficacy when applied on its own in phase III
clinical studies, was not approved due to several unacceptable
side effects, including renalꢀrelated adverse events and serum
creatinine elevations.15
intrigued to further study the two atropisomers of lesinurad in
details.
Herein, we report the discovery, characterization and assessꢀ
ment of (+)ꢀlesinurad and (ꢀ)ꢀlesinurad (Scheme 1) on their
stabilities, physicochemical properties, in vitro and in vivo
ADME/PKs. To the best of our knowledge, the existence of
atropisomers in lesinurad was not previously described.
Scheme 1. Discovery of Atropisomers of Lesinurad
The chemical structure of lesinurad was first reported in 2006
and was composed of a naphthalene ring linked with a triaꢀ
zole.16,17 Based on the structure, the triazole ring should rotate
freely along the CꢀN bond, however, the thioꢀacetic acid moieꢀ
ty and bromine atom might hinder the free rotation due to steꢀ
ric bulk. This gave us a hint that the less noticed axial chirality
possibly existed in lesinurad. A quick chiral supercritical fluid
chromatography (SFC) analysis of lesinurad confirmed our
suspection and two identical atropisomeric peaks were obꢀ
served. Considering that atropisomers might have different
pharmacological and pharmacokinetic properties, we were
Atropisomers are conformational isomers, which epimerize
or racemize via rotation along the bond axis.18 Atropisomeric
compounds represent an interesting but not fully studied famiꢀ
ly despite its importance to drug discovery and developꢀ
ment.19,20 A recent review from LaPlante and coworkers proꢀ
posed that atropisomers could be classified into three groups
based on rotational energy barriers and racemization rates (t1/2)
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(Class 1: rapid equilibration with t1/2 less than minutes; Class 2: and hERG inhibitions. The two atropisomers showed compaꢀ
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4
moderate rate of equilibration with t1/2 from hours to days;
Class 3: very slow equilibration with t1/2 in years).18,21 Since
atropisomers in Class 3 are as stable as classical chiral comꢀ
pounds, they could be isolated and treated as single enantioꢀ
mers.
rable physiochemical and biological properties in these in vitro
tests (Table 1).
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In order to find out whether the two atropisomers of lesinurad
could easily interconvert under various conditions, chiral SFC
separation was tried on either lesinurad itself or lesinuradꢀester
1 as shown in Scheme 1 and (+)/(ꢀ)ꢀlesinurad were obtained in
good yields. The Xꢀray crystal structures of both compound
1A and 1B were obtained (Figure 1) allowing confirmation of
the absolute configurations of both (+)/(ꢀ)ꢀlesinurad.
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The stability studies of (+)/(ꢀ)ꢀlesinurad in solid form, in solꢀ
vent and in plasma were performed. It was found that these
two atropisomers were very stable in their solid states and no
significant purity reduction or racemization was observed after
storage at ambient temperature for over 80 days. (+)/(ꢀ)ꢀ
lesinurad was also found to be very stable in solutions (ethaꢀ
nol/37 oC and DMSO/80 oC for 4 days), and no interꢀ
conversion was observed in these two solvents under testing
conditions, even after heating in DMSO at 120 oC for 24 hours.
Additionally, stabilities of (+)/(ꢀ)ꢀlesinurad in plasma were
also measured, and both isomers remained unchanged after
o
o
incubation in human plasma/37 C or SDꢀrat plasma/37 C up
to 2 hours.
Table 1. Physiochemical and Biological Properties Compari-
son of (+)/(-)-Lesinurad with (±)-Lesinurad
Figure 1. X-ray structures of compound 1A and 1B
Physiochemical and Biologꢀ (±)ꢀ
(ꢀ)ꢀ
(+)ꢀ
These two atropisomers were stable in vitro in human plasma
and SDꢀrat plasma, and also seemed quite stable in human in
vitro microsome stability studies. In order to find out whether
interconversion could happen in vivo and whether (+)/(ꢀ)ꢀ
lesinurad could behave differently in vivo, pharmacokinetic
studies of these two atropisomers were carried out in animals.
In the first study, rats in two groups were administrated with
(+)ꢀlesinurad & (ꢀ)ꢀlesinurad respectively and plasma samples
were analyzed using chiral column detecting (+)ꢀlesinurad and
(ꢀ)ꢀlesinurad simultaneously (Table 2). Following a single IV
or PO dose of (+)ꢀlesinurad, the other isomer (ꢀ)ꢀlesinurad was
not detected in all plasma samples at different time points.
Similarly, (+)ꢀlesinurad was not detected after a single IV or
PO dose of (ꢀ)ꢀlesinurad neither. This result confirmed that
there was no enzymeꢀcatalyzed in vivo interconversion beꢀ
tween these two atropisomers in rats.
ical properties
lesinurad lesinurad lesinurad
hURAT1 IC50 (µM)
9.6±1.7a
17.3
15.1±3.1b 4.4±1.0b
1A2
2C9
2C19
2D6
3A4
39.3
78.7
10.6
92.1
32.4
>30
21.9
31.2
10.8
>100
31
>100
61.7
CYP IC50 (µM)
hERG IC50 (µM)
32.6
74.4
>30
>30
Human
Rat
<9.5
<9.5
<17.3
<38.0
<13.8
<13.0
1.29
1.84
<9.5
<17.3
<38.0
<13.8
18.7
1.26
1.62
<17.3
<38.0
<13.8
MMS
(mL/min/kg)
Mouse
Dog
Table 2. Rat PKa Profile of (+)-Lesinurad and (-)-Lesinurad
Monkey <13.0
(ꢀ)ꢀ
lesinurad
(+)ꢀ
lesinurad
(ꢀ)/(+)ꢀ
Index
AꢀB
BꢀA
1.34
1.98
Parameters
Doseb (mg/kg)
MDR1ꢀMDCK
Papp(10ꢀ6 cm/s)
2
2
an=17; bn=5
IV
AUC0ꢀinf (µM.h)
CLp (mL/min/kg)
Dosec (mg/kg)
14.4±1.5
5.8±0.6
10
15.0±1.0
5.5±0.4
10
1.0
1.1
Lesinurad was reported to inhibit hURAT1 transporter with
IC50 value of 7.3 µM in hURAT1 highly expressed HEK293
cell lines.15 The same assay was applied to evaluate the inhibiꢀ
tion activities of the two atropisomers. After multiple tests
(n=>7), the IC50 values for (+)/(ꢀ)/(±)ꢀlesinurad inhibition of
[14C]ꢀuric acid uptake by hURAT1 were determined to be 4.4
µM, 15.1 µM and 9.6 µM, respectively (Table 1). (+)ꢀ
lesinurad showed about 3 fold boost in potency as comparison
to (ꢀ)ꢀlesinurad and (±)ꢀlesinurad. The two atropisomers toꢀ
gether with lesinurad were also evaluated for permeability, in
vitro microsome stability, cytochrome P450 enzyme isoform
PO
AUC0ꢀinf (µM.h)
46.9±11.5 32.7±13.4
1.4
aRat PK data is mean±SD, n=3; bIV formulation: 1 mg/mL in
water, pH=8, nearly clear solution with few particles before filter;
cPO formulation: 2 mg/mL in water, pH=8, nearly clear solution
with few particles
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By closer examination of the PK data, it was found that the
tion were 1.4 & 1.0 fold for IV dosing group and 2.2 & 2.5
fold for PO dosing group as shown in Table 5. Although
smaller ratios were observed in the PO group compared to the
coꢀdosing study, (ꢀ)ꢀlesinurad still demonstrated significant
higher plasma exposure and urine concentration than (+)ꢀ
lesinurad in monkeys.
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two atropisomers showed similar apparent clearance and
plasma exposure in the IV dosing groups. However, (ꢀ)ꢀ
lesinurad afforded about 1.2 fold higher plasma exposure than
(+)ꢀlesinurad upon PO dosing. This observation indicated
some potential in vivo difference between the two isomers,
although the result could also arise from experimental deviaꢀ
tions.
Table 4. Monkey PKa Profile of (+)/(-)-Lesinurad after a
single dosing of (±)-Lesinurad
To make a more informative comparison, a second study was
carried out to measure the plasma concentration of (+)ꢀ
lesinurad and (ꢀ)ꢀlesinurad following a single IV dose of raꢀ
cemic (±)ꢀlesinurad at 2 mg/kg or oral administration at 10
mg/kg (Table 3). Interestingly, the concentration of (ꢀ)ꢀ
lesinurad was always 10~20% higher than that of (+)ꢀlesinurad
in all plasma samples collected at different time points. As a
result, ratios of exposure levels between (ꢀ)ꢀ and (+)ꢀlesinurad
were 1.1 and 1.2 in IV and PO groups respectively. The in
vivo differentiation could be rationalized by slight differences
in the absorption and metabolism of the two isomers, which
were not distinguishable in in vitro experiments.
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(ꢀ)ꢀ leꢀ
sinurad
(+)ꢀ
lesinurad
(ꢀ)/(+)ꢀ
Index
Parameters
Doseb
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2 mg/kg of (±)ꢀlesinurad
AUC0ꢀinf (µM.h)
CLp (mL/min/kg)
C0ꢀ24h,urine (µM)
Dosec
14.5±3.3
5.9±1.5
5.4±0.9
10.0±2.8
8.8±2.7
2.8±1.4
1.4
0.7
1.9
IV
10 mg/kg of (±)ꢀlesinurad
99.4±29.7 28.2±19.2 3.5
153.3±93.5 38.0±20.7 4.0
PO AUC0ꢀinf (µM.h)
Table 3. Rat PKa Profile of (+)/(-)-Lesinurad after a single
dosing of (±)-Lesinurad
C0ꢀ24h,urine (µM)
aMonkey PK data is mean±SD, n=3; bIV formulation: 2.00
mg/mL in DMSO/PEG400/H2O(5/40/55), clear solution; cPO
formulation: 2 mg/mL in DMSO/0.5%MC(5/95), suspension
(ꢀ)ꢀ
lesinurad
(+)ꢀ
lesinurad
(ꢀ)/(+)ꢀ
Index
Parameters
Doseb
AUC0ꢀinf (µM.h)
CLp (mL/min/kg) 9.0±2.1
2 mg/kg of (±)ꢀlesinurad
IV
9.5±2.5
8.9±1.6
9.4±1.5
1.1
1.0
Dosec
10 mg/kg of (±)ꢀlesinurad
41.9±10.2 35.6±8.0
PO
AUC0ꢀinf (µM.h)
1.2
aRat PK data is mean±SD, n=3; bIV formulation: 1 mg/mL in
water, pH=8ꢀ9, clear solution; cPO formulation: 2 mg/mL in waꢀ
ter, pH=8ꢀ9, clear solution
With this interesting finding, we further examined the pharꢀ
macokinetic behavior of (+)ꢀlesinurad and (ꢀ)ꢀlesinurad in
large animals. Following a single IV and PO dose of racemic
lesinurad to male Cynomolgus monkeys, the two isomers were
analyzed separately. In the third PK study, the urine samples
collected during different time periods were also analyzed
since around 20% of (±)ꢀlesinurad was reported to be excreted
unchanged to urine and thus gave efficacy.15 Surprisingly,
concentrations of (ꢀ)ꢀlesinurad in both plasma and urine at all
time points/periods were found to be significantly higher than
those of (+)ꢀlesinurad in both IV and PO dosing groups (Table
4). The plasma concentrationꢀtime curves of (+)ꢀlesinurad and
(ꢀ)ꢀlesinurad are shown in Figure 2. In the IV dosing group,
the apparent clearance of (ꢀ)ꢀlesinurad was lower than that of
(+)ꢀlesinurad resulting in a 1.4 fold higher plasma exposure for
(ꢀ)ꢀlesinurad. The mean urine concentration of (ꢀ)ꢀlesinurad
was also 1.9 fold higher than that of (+)ꢀlesinurad. In the PO
dosing group, the ratios of (ꢀ)ꢀlesinurad vs (+)ꢀlesinurad for
plasma exposure and mean urine concentration were 3.5 and
4.0 fold. Taken together, these results strongly suggested that
these two atropisomers were absorbed, metabolized and exꢀ
creted differently in monkeys.
Figure 2. Plasma Concentration-Time Curve of (+)/(-)-
Lesinurad in Monkey PK Study
Table 5. Monkey PKa Profile of (+)-Lesinurad and (-)-
Lesinurad
(ꢀ)ꢀ
lesinurad
(+)ꢀ
lesinurad
(ꢀ)/(+)ꢀ
Index
Parameters
Doseb (mg/kg)
2
2
AUC0ꢀinf (µM.h)
CLp (mL/min/kg)
26.7±8.8
19.2±7.3
1.4
IV
3.3±1.0
4.7±1.5
0.7
1.0
C0ꢀ24h,urine (µM)
Dosec (mg/kg)
16.3±7.4
10
15.7±3.3
10
PO AUC0ꢀinf (µM.h)
C0ꢀ24h,urine (µM)
83.9±12.1
37.3±6.9
2.2
2.5
120.1±82.7 47.8±20.3
aMonkey PK data is mean±SD, n=3; bIV formulation: 2.00
mg/mL in DMSO/PEG400/H2O(5/40/55), clear solution; cPO
formulation: 2 mg/mL in DMSO/0.5%MC(5/95), suspension
Considering the potential drugꢀdrugꢀinteractions between the
two isomers in absorption, metabolism and excretion, we carꢀ
ried out a fourth pharmacokinetic study dosing them separateꢀ
ly in monkeys. As it turned out, the ratios of (ꢀ)ꢀlesinurad vs
(+)ꢀlesinurad for plasma exposure and mean urine concentraꢀ
Based on the animal pharmacokinetic data, (ꢀ)ꢀlesinurad
showed much higher plasma/urine exposures than (+)ꢀ
lesinurad, especially in monkeys. It was reasonable to suspect
that the two atropisomers behave differently in human. Acꢀ
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cording to the NDA reports of lesinurad, human liver cytoꢀ bromoacetyl chloride
Page 4 of 6
provided (R)ꢀ1ꢀphenylethylꢀ2ꢀ
3
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chrome P450 isozyme CYP2C9 played a major role in the
formation of oxidative metabolites while other CYPs contribꢀ
uted minimally for the metabolism of lesinurad in vitro.15 In
order to predict which atropisomer would be more stable in
human, we evaluated the stabilities of (+)ꢀlesinurad and (ꢀ)ꢀ
lesinurad in pooled human recombinant CYP2C9. Interestingꢀ
ly, it was found that (ꢀ)ꢀlesinurad was metabolized faster by
CYP2C9 with t1/2 around 90.3 min (Table 6) while (+)ꢀ
lesinurad showed prolonged t1/2 over 145 min. From the
CYP2C9 stability data, it was anticipated that (+)ꢀlesinurad
might be metabolically more stable in human than (ꢀ)ꢀ
lesinurad and might give higher exposure in plasma and urine.
This would be different from what we’d observed in animals.
As described early in this paper, (+)ꢀlesinurad was also identiꢀ
fied to be about 3 fold more potent than (ꢀ)ꢀlesinurad. Thereꢀ
fore, it was suspected that (+)ꢀlesinurad might demonstrate
better clinical efficacy than (ꢀ)ꢀlesinurad and (±)ꢀlesinurad.
Moreover, from a MetꢀID of the two isomers, three monoꢀ
oxidative metabolites (P+16; m/z 420) were detected for (ꢀ)ꢀ
lesinurad while only one monoꢀoxidative metabolite (P+16;
m/z 420) was detected for (+)ꢀlesinurad. It has been reported
that the clinical side effects observed in lesinurad might be
attributed to the oxidative metabolites.15 Taken together, (+)ꢀ
lesinurad might demonstrate a larger therapeutic index than (ꢀ
)ꢀlesinurad or the racemate.
bromoacetate 4. Sꢀalkylation of thioꢀtrizole 5 gave intermediꢀ
ate 6. Further bromination yielded key intermediate 7, and 7A
& 7B could be separated by column. Subsequent hydrolysis of
7A and 7B gave (+)ꢀlesinurad and (ꢀ)ꢀlesinurad in high yields.
Interestingly, we did not observe the existence of stable atroꢀ
pisomers in compound 6 under various SFC conditions, which
indicated the observed axial chirality of (+)ꢀlesinurad and (ꢀ)ꢀ
lesinurad was maintained by the introduction of steric bulky
bromine atom.
9
In conclusion, existence of stable atropisomers was discovꢀ
ered in lesinurad. (+)ꢀ/(ꢀ)ꢀlesinurad were isolated and fully
characterized. No interconversion was observed between the
two isomers under heat, in solutions and in in vivo pharmacoꢀ
kinetic studies. Although the two atropisomers showed compaꢀ
rable data in most of the physiochemical properties, (+)ꢀ
lesinurad showed significant higher in vitro inhibition potency
against hURAT1 than (ꢀ)ꢀlesinurad. Furthermore, the two atꢀ
ropisomers showed significantly different pharmacokinetic
profiles in Cynomolgus monkeys. In vitro human recombinant
CYP2C9 stability study indicated that (+)ꢀlesinurad was more
stable than (ꢀ)ꢀlesinurad. Considering the significant difference
between the two isomers, it was speculated that the (+)ꢀ
lesinurad might offer a better hyperuricemia/gout therapy than
(ꢀ)ꢀlesinurad or the racemate.
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ASSOCIATED CONTENT
Supporting Information
Table 6. Microsome Stability of (+)/(-)-Lesinurad in Re-
combinant Human CYP2C9
a
b
Compound
(ꢀ)ꢀlesinurad
(+)ꢀlesinurad
Diclofenac
t1/2
CLint
0.04
Stability 1c Stability 2d
The Supporting Information is available free of charge on the
ACS Publications website.
90.3
61.8%
74.9%
3.3%
110.5%
108.9%
112.2%
>145.0
<0.02
Full experimental details, in vitro and in vivo assay. (PDF)
12.3
5.6
aValue was reported in min.; bValue was reported in
AUTHOR INFORMATION
ꢁL/min/pmol; cValue was reported as the %remaining at T=60
min in the presence of coꢀfactor NADPH.; dValue was reported as
the %remaining at T=60 min in the absence of coꢀfactor NADPH.
Corresponding Author
* Eꢀmail: Zhang_yang@wuxiapptec.com for Dr Yang Zhang.
Scheme 2. Chiral Synthesis of (+)/(-)-Lesinurad
Author Contributions
The manuscript was written through contributions of all authors.
All authors have given approval to the final version of the manuꢀ
script.
ACKNOWLEDGMENT
We acknowledge Dr. Yikai Wang for the helpful discussions and
review of this manuscript.
ABBREVIATIONS
hURAT1, human uric acid transport 1; PK, pharmacokinetics;
CYP, Cytochrome P450; hERG, human etherꢀaꢀgoꢀgo related
gene; MMS, microsomes metabolic stability; MetꢀID, metabolism
identification; IV, intravenous; PO, per oral; AUC, area under
curve; CLp, plasma clearance; CLint, intrinsic clearance; SD rat,
SpragueꢀDawley Rat; NADPH, βꢀNicotinamide adenine dinucleoꢀ
tide phosphate;
REFERENCES
(1) Liu, B.; Wang, T.; Zhao, H. N.; Yue, W. W.; Yu, H. P.; Liu, C.
X.; Yin, J.; Jia, R. Y.; Nie, H. W. The prevalence of hyperuricemia in
China: a metaꢀanalysis. BMC Public Health 2011, 11, 832.
Next, with the effort to develop a readily accessible way to
prepare (+)ꢀlesinurad and (ꢀ)ꢀlesinurad without costly SFC
separation, a chiral synthetic route was developed as described
in the Scheme 2. Acylation of (R)ꢀ1ꢀphenylethanol 2 with 2ꢀ
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(2) Zhu, Y.; Pandya, B. J.; Choi, H. K. Prevalence of gout and
concentrative nucleoside transporter 2 inhibitors through suppressing
intestinal absorption of purine nucleosides. Eur. J. Pharmacol. 2012,
690, 183ꢀ191.
(13) Yeh, L.; Shen, Z.; Kerr, B. RDEA594: a potent URAT1 inhibꢀ
itor without affecting other important renal transporters, OAT1 and
OAT 3. Ann Rheum Dis. 2009, 68, 320.
(14) Fleischmann, R.; Kerr, B.; Yeh, L. T.; Suster, M.; Shen, Z. C.;
Polvent, E.; Hingorani, V.; Quart, B.; Manhard, K.; Jeffrey, N. M.;
Baumgartner, S. Pharmacodynamic, pharmacokinetic and tolerability
evaluation of concomitant administration of lesinurad and febuxostat
in gout patients with hyperuricaemia. Rheumatology 2014, 53, 2167ꢀ
2174.
hyperuricemia in the US general population: the National Health and
Nutrition Examination Survey 2007ꢀ2008. Arthritis Rheum. 2011, 63,
3136ꢀ3141.
1
2
3
4
5
6
7
8
(3) DeOliveira, E. P.; Burini, R. C. High plasma uric acid concenꢀ
tration: causes and consequences. Diabetol Metab Syndr. 2012, 4, 12.
(4) Zhang, W.; Doherty, M.; Pascual, E. EULAR evidence based
recommendations for gout. Part I: Diagnosis. Report of a task force of
the Standing Committee for International Clinical Studies Including
Therapeutics (ESCISIT). Ann Rheum Dis. 2006, 65, 1301ꢀ1311.
(5) Zhang, W,; Doherty, M.; Bardin, T. EULAR evidence based
recommendations for gout. Part II: Management. Report of a task
force of the EULAR Standing Committee for International Clinical
Studies Including Therapeutics (ESCISIT). Ann Rheum Dis. 2006, 65,
1312ꢀ1324.
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
(15) Center for Drug Evaluation and Research. Clinical Pharmaꢀ
cology
and
Biopharmaceutics
Review(s).
http://www.accessdata.fda.gov/drugsatfda_docs/nda/2015/207988Ori
g1s000ClinPharmR.pdf. Jan. 19, 2016
(16) Pema, K. M. Lesinurad sodium. Solute carrier family 22
member 12 (URAT1) inhibitor, uricosuric, treatment of gout. Drugs
of the Future 2011, 36, 875ꢀ880.
(17) Girardet, J. L.; KOH, Y. H.; Delarosa, M.; Gunic, E.; Hong,
Z.; Lang, S.; Kim, H. W. WO 2006/026356.
(18) LaPlante, S. R.; Fader, L. D.; Fandrick, K. R.; Fandrick, D. R.;
Hucke, O.; Kemper, R.; Miller, S. P. F.; Edwards, P. J. Assessing
Atropisomer Axial Chirality in Drug Discovery and Development. J.
Med. Chem. 2011, 54, 7005ꢀ7022.
(19) Bringmann, G.; Mortimer, A. J. P.; Keller, P. A.; Gresser, M.
J.; Garner, J.; Breuning, M. Atroposelective Synthesis of Axially
Chiral Biaryl Compounds. Angew. Chem. Int. Ed. 2005, 44, 5384ꢀ
5427.
(20) Maryanoff, B. E.; Greco, M. N. Stereochemical Lability in
Drug Molecules: Cases Where Chirality May Not Be Critical for
Drug Development. In Comprehensive Chirality Vol. 1, Carriera, E.
M., Yamamto, H., Eds.; Elsevier, Amsterdam, NL, 2012; pp 105ꢀ119.
(21) LaPlante, S. R.; Edwards, P. J.; Fader, L. D.; Jakalian, A.;
Hucke, O. Revealing atropisomer axial chirality in drug discovery.
ChemMedChem 2011, 6, 505ꢀ513.
(6) Daria, B. C.; Michael, H. P. New Therapies for Gout. Annu.
Rev. Med. 2013, 64, 325ꢀ337.
(7) Hania, S.; Jasvinder, A. S. Investigational drugs for hyperuꢀ
ricemia. Expert Opin. Investig. Drugs. 2015, 24, 1013ꢀ1030.
(8) Shoji, A.; Yamanaka, H.; Kamatani, N. A retrospective study of
the relationship between serum urate level and recurrent attacks of
gouty arthritis: evidence for reduction of recurrent gouty arthritis with
ntihyperuricemic therapy. Arthritis Rheum. 2004, 51, 321ꢀ325.
(9) Cesar, D. T.; Nuria, P. H.; Fernando P. R. New medications in
development for the treatment of hyperuricemia of gout. Curr Opin
Rheumatol. 2015, 27, 164ꢀ169.
(10) Matsuo, H.; Nakayama, A.; Sakiyama, M. ABCG2 dysfuncꢀ
tion causes hyperuricemia due to both renal urate underexcretion and
renal urate overload. Scientific Reports 2014, 4, 3755ꢀ3799.
(11) Ahn, S. O.; Ohtomo, S.; Kiyokawa, J.; Nakagawa, T.; Yamaꢀ
ne,M.; Lee, K. J.; Kim, K. H.; Kim, B. H.; Tanaka, J.; Kawabe, Y.;
Horiba, N. Stronger uricosuric effects of the novel selective URAT1
inhibitor URꢀ1102 lowered plasma urate in tufted capuchin monkeys
to a greater extent than benzbromarone. Journal of Pharmacology &
Experimental Therapeutics 2016, 357, 157ꢀ166.
(12) Hiratochi, M.; Tatani, K.; Shimizu, K.; Kuramochi, Y.; Kikuꢀ
chi, N.; Kamada, N.; Itoh, F.; Isaji, M. Hypouricemic effects of novel
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