5-Substituted-N-pyridazinylbenzamides as potent and selective LRRK2 inhibitors: Improved brain unbound fraction enables efficacy

Article abstract of DOI:10.1016/j.bmcl.2018.11.054

We describe the discovery and optimization of 5-substituted-N-pyridazinylbenzamide derivatives as potent and selective LRRK2 inhibitors. Extensive SAR studies led to the identification of compounds 18 and 23, which demonstrated good in vitro pharmacokinetic profile and excellent selectivity over 140 other kinases. Both compounds demonstrated high unbound fractions in both blood and brain. Compound 18 proved to be brain penetrant, and the high unbound fraction of compound 18 in brain enabled its in vivo efficacy in CNS, wherein a significant inhibition of LRRK2 Ser935 phosphorylation was observed in rat brain following intravenous infusion at 5 mg/kg/h.

Full text of DOI:10.1016/j.bmcl.2018.11.054

Accepted Manuscript
5-Substituted-N-pyridazinylbenzamides as Potent and Selective LRRK2 Inhib-
itors: Improved Brain Unbound Fraction Enables Efficacy
Xiao Ding, Luigi Piero Stasi, Xuedong Dai, Kai Long, Cheng Peng, Baowei
Zhao, Hailong Wang, Changhui Sun, Huan Hu, Zehong Wan, Karamjit S. Jandu,
Oliver J. Philps, Yan Chen, Lizhen Wang, Qian Liu, Colin Edge, Yi Li, Kelly
Dong, Xiaoming Guan, F. David Tattersall, Alastair D. Reith, Feng Ren
PII:
S0960-894X(18)30930-2
https://doi.org/10.1016/j.bmcl.2018.11.054
BMCL 26165
DOI:
Reference:
Bioorganic & Medicinal Chemistry Letters
To appear in:
Received Date:
Revised Date:
Accepted Date:
13 September 2018
24 November 2018
27 November 2018
Please cite this article as: Ding, X., Stasi, L.P., Dai, X., Long, K., Peng, C., Zhao, B., Wang, H., Sun, C., Hu, H.,
Wan, Z., Jandu, K.S., Philps, O.J., Chen, Y., Wang, L., Liu, Q., Edge, C., Li, Y., Dong, K., Guan, X., Tattersall,
F.D., Reith, A.D., Ren, F., 5-Substituted-N-pyridazinylbenzamides as Potent and Selective LRRK2 Inhibitors:
Improved Brain Unbound Fraction Enables Efficacy, Bioorganic & Medicinal Chemistry Letters (2018), doi: https://
doi.org/10.1016/j.bmcl.2018.11.054
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers
we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and
review of the resulting proof before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Graphical Abstract
To create your abstract, type over the instructions in the template box below.
Fonts or abstract dimensions should not be changed or altered.
5-Substituted-N-pyridazinylbenzamides as Potent and
Selective LRRK2 Inhibitors: Improved Brain Unbound
Fraction Enables Efficacy
Leave this area blank for abstract info.
Xiao Dinga, Luigi Piero Stasia, Xuedong Daia, Kai Longa, Cheng Penga, Baowei Zhaoa, Hailong Wanga,
Changhui Suna, Huan Hua, Zehong Wana, Karamjit S. Jandub, Oliver J. Philpsb, Yan Chena, Lizhen Wanga, Qian
Liuc, Colin Edged, Yi Lic, Kelly Dongc, Xiaoming Guana, F. David Tattersalla, Alastair D. Reithb, Feng Ren*, a

Bioorganic & Medicinal Chemistry Letters
5-Substituted-N-pyridazinylbenzamides as Potent and Selective LRRK2
Inhibitors: Improved Brain Unbound Fraction Enables Efficacy
Xiao Dinga, Luigi Piero Stasia, Xuedong Daia, Kai Longa, Cheng Penga, Baowei Zhaoa, Hailong Wanga, Changhui
Suna, Huan Hua, Zehong Wana, Karamjit S. Jandub, Oliver J. Philpsb, Yan Chena, Lizhen Wanga, Qian Liuc, Colin
Edged, Yi Lic, Kelly Dongc, Xiaoming Guana, F. David Tattersalla, Alastair D. Reithb, Feng Ren*, a
aNeurodegeneration DPU, Neurosciences Therapeutic Area Unit, GSK Pharmaceuticals R&D, 898 Halei Road, Zhangjiang Hi-Tech Park, Pudong, Shanghai
201203, PR China.
bNeurodegeneration DPU, Neurosciences Therapeutic Area Unit, GSK Pharmaceuticals R&D, Medicines Research Centre, Gunnels Wood Road, Stevenage,
Herts, UK.
cPlatform Technology Sciences, GSK Pharmaceuticals R&D, 898 Halei Road, Zhangjiang Hi-Tech Park, Pudong, Shanghai 201203, PR China.
dPlatform Technology Sciences, GSK Pharmaceuticals R&D, Medicines Research Centre, Gunnels Wood Road, Stevenage, Herts UK.
*Corresponding author. Tel.: +86 18616743377; fax: +86 021 8596369; e-mail address: 2648666968@qq.com (F. Ren).
ARTICLE INFO
ABSTRACT
Article history:
Received
Revised
Accepted
Available online
We describe the discovery and optimization of 5-substituted-N-pyridazinylbenzamide
derivatives as potent and selective LRRK2 inhibitors. Extensive SAR studies led to the
identification of compounds 18 and 23, which demonstrated good in vitro pharmacokinetic
profile and excellent selectivity over 140 other kinases. Both compounds demonstrated high
unbound fractions in both blood and brain. Compound 18 proved to be brain penetrant, and the
high unbound fraction of compound 18 in brain enabled its in vivo efficacy in CNS, wherein a
significant inhibition of LRRK2 Ser935 phosphorylation was observed in rat brain following
intravenous infusion at 5 mg/kg/h.
Keywords:
CNS penetration
LRRK2
Parkinson’s disease
PFI
2009 Elsevier Ltd. All rights reserved.
unbound fraction
Parkinson’s disease (PD) is a common and complex
neurological disorder which affects approximately 2-3% of the
population greater than 60 years of age1-3. Currently available
treatments can only address the dopamine deficiencies to
relieve the symptoms4, and a disease modifying therapy to halt
the progression of disease remains a major unmet medical need
5. Over the past several years, great progress has been made in
the field of PD genetics and several common variants have
been identified by genome-wide association studies (GWAS)6.
Among them, mutations of Leucine-rich repeat kinase 2
(LRRK2) genes have been identified as the most common
known genetic cause of PD, accounting for 4% of familial
cases and 1% of sporadic cases7-10. Several LRRK2 mutations,
with the most frequent one G2019S located in the activation
loop of the kinase domain10-11, have been demonstrated to
increase kinase activity both in vitro and in vivo. Hence, small-
molecules that inhibit LRRK2 kinase activity could be a new
class of disease-modifying agents for the treatment of PD.
Several structurally diverse LRRK2 inhibitors have been
reported12-13 in the literature including 7H-pyrrolo[2,3-
d]pyrimidin-2-amines14-15, indazoles16-17, indolinones18 and 2,4-
diaminopyrimidines19.
We recently reported the discovery of a series of 5-
substituent-N-arylbenzamide derivatives as LRRK2 inhibitors
exemplified by 1 and 2 (Scheme 1)20-21. Given the structural
novelty, high potency and good selectivity across the kinome,
compound 1 (GSK2578215A) has been widely used as a tool
for the exploration of biological roles of LRRK222-23. However,
despite its high exposure in brain, no significant inhibition
against phosphorylation of Serine 935 (Ser935) of LRRK2 was
observed. We hypothesized its extremely low unbound fraction
in brain could be accountable for its lack of target engagement
in CNS. Besides, both compounds 1 and 2 showed poor
developability profile (e.g. solubility). Herein, we describe the
optimization starting from compound
2
towards the
identification of 5-substituted-N-pyridazinylbenzamides as
potent and selective LRRK2 kinase inhibitors with improved
developability profile. Representative compounds 18 and 23
demonstrated good in vitro pharmacokinetic profile and
excellent kinase selectivity. More importantly, these
compounds exhibited much improved free fractions in both
blood and brain in rodent species as well as in human serum.
The high brain unbound fraction of compound 18 enabled its in
vivo efficacy, wherein a significant inhibition of Ser935

phosphorylation of LRRK2 was observed in rat brain
following intravenous infusion at 5 mg/kg/h.
and 12). A more significant improvement in metabolic stability
was observed for the 6-membered ring analogue 14. Further
reduction of the ChromLogD7.4 by introducing the hydroxy
group resulted in compounds (15 and 16) with excellent
metabolic stability. However, both compounds started to show
Pgp/BCRP efflux liability with permeability ratio (PR) > 2.0,
raising concerns for CNS penetration. The Pgp/BCRP efflux of
both compounds might be due to the additional hydrogen bond
donor from the hydroxy group. Changing the hydroxy group
to the cyano group (17) mitigated the Pgp/BCRP efflux
liability, whereas the metabolic stability was still above our
cut-off value of 3 mL/min/g thus was not acceptable. A
balanced profile was observed for the morpholine analogue 18,
which demonstrated acceptable metabolic stability (2.1
mL/min/g) in human liver microsome, free of Pgp/BCRP
efflux liability (PR = 1.3), good permeability (500 nm/s), and
decent solubility (68 M). Introducing of methyl group(s) to
the morpholine moiety didn’t further improve the overall
profile.
Figure 1. Structures of compounds 1 and 2
Our work was initiated with re-evaluation of different
substitutions on the N-aryl group (R1) and benzyl ether moiety
(R2) of compound 2 to improve physicochemical properties
and developability profile. ChromlogD7.4 and property forecast
index (PFI)24 served as key indicators of developability since
they have been reported to show good relationship with
hydrophobicity,
solubility
and
other
developability
parameters24. As described previously20, compound 2 displayed
high potency against LRRK2 inhibition (HTRF pIC50 = 7.7),
good in vitro and in vivo pharmacokinetic profile but high
tissue binding in mouse brain which was proposed to be
responsible for the lack of in vivo efficacy. Further profiling
revealed low solubility (11 µM), high ChromlogD7.4 (6.3) and
PFI (9.3) of the compound. Previous SAR showed only limited
substituents could be tolerated at R1 position since this group
was proposed to be buried into the conservative pockets close
to the gatekeeper20. As illustrated in Table 1, replacing the
pyridine group (2 and 3) with 3-chlolo-phenyl (4 and 5) or 3-
fluoro-phenyl (6) substituents on R1 resulted in similar
potencies but around 2 units higher of PFI values. The
corresponding nitrile analogue (7) demonstrated increased
potency (pIC50= 8.1) and lower PFI (10.5) compared to that of
the fluoro analogue (6). Compounds with 4-pyridazinyl
substituent at the R1 position gave the improved PFI (8.4) and
comparable potency as compound 2. However, no
improvement in solubility was observed for compound 8 even
though the lipophilicity and PFI were reduced. Nevertheless,
the 4-pyridazinyl group was identified as the optimal R1
moiety in terms of the balance of potency and PFI.
Table 1. LRRK2 inhibition activities for benzamide
compounds 2‒8.
HTRF
pIC50
ChromlogDa
(PFI) b
Solubilityc
(uM)
Cpd
R1
R2
2
3
F
7.7
7.9
6.3 (9.3)
6.3 (9.3)
11
<1
H
4
5
6
F
H
F
7.9
7.6
7.5
8.7 (11.7)
8.8 (11.8)
8.2 (11.2)
NDd
7
<1
We then focused our optimization on exploring substituents
on the 5-position of the benzamide (R3), aiming to further
reduce PFI for improved solubility and developability profile
(Table 2). A variety of different groups were synthesized and
their potencies, physicochemical properties and in vitro
pharmacokinetic profile were determined. Not surprisingly, the
HRTF potency against LRRK2 inhibition for these compounds
was retained with O-, N- and C-linked substituents, consistent
with the previous predicted binding mode that R3 group was
pointed to the solvent-exposed area of LRRK2 kinase20.
Replacing the high lipophilic group CF3 with ether group such
as methoxy (9) gave a comparable potency (pIC50 = 7.8) and
much reduced lipophilicity. However, O-linked compounds
suffered from high human liver microsome clearance24 which
was probably due to the oxidative de-alkylation of the alkoxy
group. Changing R3 to N-linked moieties, such as cyclic-amine
substituents with different ring size (compound 1026)
provided improved PFI values compared with compound 8
except for compounds 12 and 13. As a result, the solubility of
these compounds was improved significantly with compounds
10 and 17 as exceptions. Compared with the O-linked analogue
9, the metabolic stabilities were slightly improved for the 4-
membered and 5-membered ring analogues (compounds 10, 11,
7
8
F
8.1
7.8
7.5 (10.5)
5.4 (8.4)
<1
<1
H
aMeasured ChromLogDpH7.4 bProperty Forecast Index = ChromLog
.
DpH7.4 + # Ar. cKinetic solubility. dNot determined.
We then started to explore the piperazine substituents, and
all piperazine derivatives (2123) demonstrated good HTRF
potency against LRRK2 inhibition. Not surprisingly, with one
more basic center, the ChromlogD7.4 and PFI values were
reduced and solubility was increased. In addition, the
metabolic stabilities of these analogues were even better than
the morpholine analogues (18 and 20), which could be partially
attributed to their lower ChromLogD7.4 values. The free
piperazine analogue 21 showed significant Pgp/BCRP efflux
liability, likely attributed to its additional hydrogen bond donor.
Masking the N-H bond with a methyl group (22) mitigated the
Pgp/BCRP efflux however with the borderline PR (1.8).
Further increasing the size of the side chain led to compound
23 with no concern of Pgp/BCRP efflux (PR = 1.3).

unlikely to be progressed as CNS compounds. Further,
changing the piperazine moiety of compounds 22 and 23 to
piperidine moieties gave C-linked analogs 27 and 28 with
increased Pgp/BCRP efflux liabilities (PR = 6.9 and 2.5,
respectively), which might be due to the higher basicity of the
piperidine compared with piperazine.
The solubility of 23 was measured high (373 M) and its
clearance was measured low (1.3 mL/min/g). The fused amine
(24) and spiro-amines (25 and 26) were also evaluated.
Despite their good potencies and solubility, they suffered from
either high clearance or Pgp/BCRP efflux liability thus was
Table 2. LRRK2 inhibition activities and in vitro profile of benzamide compounds 8‒28.
Pgp/Bcrpd PRe
PP (nm/s) f
solubilityg
(uM)
hClic (mL/min/g)
Cpd
R3
HTRF pIC50
ChromLogDa (PFI)b
8
CF3
7.8
7.8
7.6
5.4 (8.4)
4.9 (7.9)
4.4 (7.4)
NDi
22.6
11.8
NDi
<1
NDi
3
9h
OCH3
NDi
10
11
12
1.3/563
7.9
7.7
5.1 (8.1)
6.1 (9.1)
9.5
9.8
1.0/390
2.4/184
29
35
13
14
15
7.6
7.4
7.4
6.3 (9.3)
5.1 (8.1)
3.2 (6.2)
NDi
5.5
0.6
NDi
60
20
28
1.1/508j
2.1/470j
16
17
18
7.7
8
3.5 (6.5)
4.5 (7.5)
4.1 (7.1)
1
4.1/300
1.1/483j
1.3/500
323
<1
5.5
2.1
7.4
68
19
20
7.7
7.7
4.5 (7.5)
5.4 (8.4)
NDi
2.2
NDi
113
29
1.5/592j
21
22
7.8
7.9
1.9 (4.9)
3.0 (6.0)
0.4
1.6
4.9/141
1.8/406
98
228
23
7.7
3.8 (6.8)
1.3
1.3/457
373
24
25
26
27
7.3
7.6
7.2
7.7
4.5 (7.5)
2.0 (5.0)
2.2 (5.2)
2.3 (5.3)
4.3
NDi
NDi
NDi
1.6/660j
7.6/426
9.9/97
57
198
266
71
6.9/183
28
7.2
2.9 (5.9)
<0.78
2.5/414
226
aMeasured ChromLogDpH7.4. bProperty Forecast Index = ChromLogDpH7.4 + # Ar. cHuman liver microsome clearance. dMDCKII-MDR1 transduced with
BacMam2-BCRP cell line. ePermeability ratio, A→B (apical to basolateral) with GF120918/A→B without GF120918. fPassive permeability, A→B
with GF120918. gKinetic solubility. hPyridyl analogue instead of pyridazinyl. iNot determined. jMDCKII-MDR1 cell line.

Ser935 phosphorylation in brain, kidney and spleen was
determined.
Table 3. Developability profile of compounds 18 and 23
18
23
a
ANK/AHE pIC50
7.0 / 6.8
1/140
13/4.6
12.5
6.6 / 6.4
1/140
11/6.4
13.6
Kinase selectivityb
Fu% in blood/brainc
Fu% in HSAd
CYP inhibition pIC50 (3A4,
atorvastatin)
5.2
5.0
hPXR pEC50
OATP1B1 pIC50
<4.3
4.8
<4.3
4.4
hERG IonWorks pIC50
<4.2
5.1
aHuman lymphoblastoid cells derived from
a
control (AHE) and
Parkinson’s patient (ANK) homozygous for the LRRK2[G2019S]
mutation. bStandard radioactivity-based enzymatic assays against a panel
of 140 kinases at 1 uM, quoted as numbers of kinase displaying >50%
activity. cFree fraction in rat brain and blood. dFree fraction in human
serum albumin.
Compound
and
were then selected for further progression
23
18
(Table 3) given their high potencies, decent physicochemical
properties (ChromlogD7.4, PFI, and solubility), good metabolic
stabilities, and Pgp/BCRP efflux profile. The in vitro
pharmacology of compounds was examined on endogenous
LRRK2 in human lymphoblastoid cells derived from a healthy
subject (AHE) and
a
Parkinson’s disease patient with
18
Figure 2.Pharmacodynamic study of 18 at 4 h after intravenous
infusion at dose 5 mg/kg/h.
homozygous LRRK2[G2019S] mutation (ANK).26 Compound
demonstrated pIC50 values of 6.8 in AHE and 7.0 in ANK cell
assays. Relatively lower potencies were observed for compound
To our delight, ~50% reduction on phosphorylation of LRRK2
Ser935 in brain was observed at 4 h after iv infusion (5 mg/kg/h)
in rats (Figure 2). In addition, ~60% and ~70% reductions on
phosphorylation of LRRK2 Ser935 were observed in kidney and
spleen, respectively. Pharmacokinetics (PK) analysis revealed
drug concentrations in blood, brain, kidney, and spleen at steady
state were 1126 ng/mL, 1627 ng/g, 3967 ng/g, 1680 ng/g,
respectively. Compound 18 proved to be brain penetrant with
good brain-to-blood ratios in both total drug concentration (Kp =
1.44) and free drug concentration (Kp,uu = 0.51). The drug
concentration of compound 18 in brain correlated well with its in
vivo CNS efficacy. This result, comparing with the previous
report of the lack of correlation between CNS efficacy and drug
exposure for compound 1, suggested that the free unbound
fraction of compound 18 in brain enabled its in vivo efficacy in
CNS.
with pIC50 values of 6.4 and 6.6 in AHE and ANK assays,
23
respectively. Both compounds were progressed to kinase
selectivity assessment using standard radioactivity-based
enzymatic assays at the concentration of 1 µM. They both
demonstrated excellent selectivity against a panel of 140 kinases
among which both compounds hit only one kinase, 84%
inhibition against mitogen-activated protein kinase kinase 1
(MKK1) for compound
and 53 % inhibition against AMPK-
18
related protein kinase 5 (ARK5) for compound . The unbound
23
fractions were also determined in rat blood and brain tissue.
Compounds
and
demonstrated good free fractions in both
23
18
blood (13% and 11%, respectively) and brain (4.6% and 6.4%,
respectively). Consistently, good unbound fractions were also
observed in human serum albumin (HSA) with 12.5% and 13.6
% for compounds
and
, respectively. Further, the
23
18
developability profile was evaluated for both compounds. They
demonstrated moderate inhibition against human CYP3A4 and
low inhibition on OATP1B1, and no meaningful activity against
human PXR (pIC50 < 4.3), indicating low concerns for drug-
drug interactions. In addition, the preliminary cardiac safety was
evaluated by conducting the hERG binding assay, and the
In conclusion, through extensive SAR studies and careful
modifications of ChromLogD7.4 and PFI starting from compound
2,
we
discovered
a
series
of
5-substituted-N-
pyridazinylbenzamide derivatives with high potencies of LRRK2
inhibition, improved physicochemical properties, and
developability profile. Compounds 18 and 23 were identified
with good in vitro PK profile and excellent selectivity over 140
other kinases. Most importantly, both compounds exhibited high
free unbound fractions both in blood and brain. Compound 18
proved to be brain penetrant with good brain-to-blood ratios in
both total drug concentration (Kp = 1.44) and free drug
concentration (Kp,uu = 0.51). In vivo pharmacology study of
compound 18 demonstrated significant inhibitions of LRRK2
Ser935 phosphorylation in brain, kidney and spleen following iv
infusion (5 mg/kg/h) in rats. We concluded that the in vivo
efficacy in CNS of compound 18 was enabled by its high free
unbound fraction in brain. The desirable in vitro and in vivo
activity and developability profile of compound 18 made it a
compound was determined to have a lower risk of QT interval
18
prolongation (hERG pIC50 < 4.2) compared with compound
23
demonstrated a
(hERG pIC50 = 5.1). In this regard, compound
18
superior developability profile than compound
.
23
As reported previously, the benzamide analogue (1) suffered
from the lack of in vivo pharmacological efficacy in CNS, which
was hypothesized to due to its low free fraction in brain (0.3%).20
Discovery of compound 18 with significantly improved free
unbound fraction in brain (4.6%) provided a good tool to test the
hypothesis. In order to ensure sufficient exposure of compound
18 in brain at steady state, an intravenous (iv) bolus followed by
intravenous infusion administration was applied in the in vivo
pharmacodynamics study, wherein the inhibition of LRRK2

good tool for the exploration of the biological roles of LRRK2 in
Parkinson’s disease.
Parker, E. M.; Stamford, A. W.; Nargund, R.; McCauley, J. A.; Miller,
M. W. J. Med. Chem. 2017, 60, 2983.
17. Ding, X.; Jin, Y.; Liu, Q.; Ren, Feng.; Sang, Y.; Stasi, L. P.; Wan, Z.;
Wang, H.; Xing, W.; Zhan, Y.; Zhao, B. WO 2017012576. 2017.
18. Troxler, T.; Greenidge, P.; Zimmermann, K.; Desrayaud, S.; Druckes, P.;
Schweizer, T.; Stauffer, D.; Rovelli, G.; Shimshek, D. R. Bioorg. Med.
Chem .Lett. 2013, 23, 4085.
Acknowledgement
We thank Mr. Liang Huang for HRMS analysis, Haihua Yu for 13C
NMR determination, Mr. Lee Collier for HTRF assay determination,
Mr. Lijun Ge, Mr. Yan Chen, and Mr. Yong Zhang for their in vivo
work for compound testing.
19. Estrada, A. A.; Chan, B. K.; Baker-Glenn, C.; Beresford, A.; Burdick, D.
J.; Chambers, M.; Chen, H.; Dominguez, S. L.; Dotson, J.; Drummond,
J.; Flagella, M.; Fuji, R.; Gill, A.; Halladay, J.; Harris, S. F.; Heffron, T.
P.; Kleinheinz, T.; Lee, D. W.; Le Pichon, C. E.; Liu, X.; Lyssikatos, J.
P.; Medhurst, A. D.; Moffat, J. G.; Nash, K.; Scearce-Levie, K.; Sheng,
Z.; Shore, D. G.; Wong, S.; Zhang, S.; Zhang, X.; Zhu, H.; Sweeney, Z.
K. J. Med. Chem. 2014, 57, 921.
Supplementary Material
Supplementary data associated with this article can be found in
the online version.
20. Ding, X.; Dai, X.; Long, K.; Peng, C.; Andreotti, D.; Bamborough, P.;
Eatherton, A. J.; Edge, C.; Jandu, K. S.; Nichols, P. L.; Philps, O. J.;
Stasi, L. P.; Wan, Z.; Xiang, J. N.; Dong, K.; Dossang, P.; Ho, M. H.; Li,
Y.; Mensah, L.; Guan, X.; Reith, A. D.; Ren, F. Bioorg. Med. Chem. Lett.
2017, 27, 4034.
Reference and Notes
1. Poewe, W.; Seppi, K.; Tanner, C. M.; Halliday, G. M.; Brundin, P.;
Volkmann, J.; Schrag, A. E.; Lang, A. E. Nat. Rev. Dis. Primers 2017, 3,
17013.
2. Pringsheim, T.; Jette, N.; Frolkis, A.; Steeves, T. D. Mov. Disord 2014,
29, 1583.
21. Reith, A. D.; Bamborough, P.; Jandu, K.; Andreotti, D.; Mensah, L.;
Dossang, P.; Choi, H. G.; Deng, X.; Zhang, J.; Alessi, D. R.; Gray, N. S.
Bioorg. Med. Chem. Lett. 2012, 22, 5625.
3. Obeso, J. A.; Stamelou, M.; Goetz, C. G.; Poewe, W.; Lang, A. E.;
Weintraub, D.; Burn, D.; Halliday, G. M.; Bezard, E.; Przedborski, S.;
Lehericy, S.; Brooks, D. J.; Rothwell, J. C.; Hallett, M.; DeLong, M. R.;
Marras, C.; Tanner, C. M.; Ross, G. W.; Langston, J. W.; Klein, C.;
Bonifati, V.; Jankovic, J.; Lozano, A. M.; Deuschl, G.; Bergman, H.;
Tolosa, E.; Rodriguez-Violante, M.; Fahn, S.; Postuma, R. B.; Berg, D.;
Marek, K.; Standaert, D. G.; Surmeier, D. J.; Olanow, C. W.; Kordower,
J. H.; Calabresi, P.; Schapira, A. H. V.; Stoessl, A. J. Mov. Disord. 2017,
32, 1264.
22. Ito, G.; Katsemonova, K.; Tonelli, F.; Lis, P.; Baptista, M. A.; Shpiro, N.;
Duddy, G.; Wilson, S.; Ho, P. W.; Ho, S. L.; Reith, A. D.; Alessi, D. R.
Biochem. J. 2016, 473, 2671.
23. Steger, M.; Tonelli, F.; Ito, G.; Davies, P.; Trost, M.; Vetter, M.;
Wachter, S.; Lorentzen, E.; Duddy, G.; Wilson, S.; Baptista, M. A.;
Fiske, B. K.; Fell, M. J.; Morrow, J. A.; Reith, A. D.; Alessi, D. R.;
Mann, M. Elife 2016, 5.
24. Young, R. J.; Green, D. V.; Luscombe, C. N.; Hill, A. P. Drug Discov.
Today 2011, 16, 822.
4. Oertel, W.; Schulz, J. B. J. Neurochem. 2016, 139, 325.
5. Kalia, L. V.; Kalia, S. K.; Lang, A. E. Mov.Disord. 2015, 30, 1442.
6. Kalinderi, K.; Bostantjopoulou, S.; Fidani, L., The genetic background of
Parkinson's disease: current progress and future prospects. Acta.
Neurol.Scand. 2016, 134, 314326.
7. Zimprich, A.; Biskup, S.; Leitner, P.; Lichtner, P.; Farrer, M.;Lincoln, S.;
Kachergus, J.; Hulihan, M.; Uitti, R. J.; Calne, D. B.; Stoessl, A. J.;
Pfeiffer, R. F.; Patenge, N.; Carbajal, I. C.; Vieregge, P.; Asmus, F.;
Muller-Myhsok, B.; Dickson, D. W.; Meitinger, T.; Strom, T. M.;
Wszolek, Z. K.; Gasser, T. Neuron 2004, 44 , 601.
25. The human biological samples were sourced ethically and their research
use was in accord with the terms of the informed consents. All studies
were conducted in accordance with the GSK Policy on the Care, Welfare
and Treatment of Laboratory Animals and were reviewed the
Institutional Animal Care and Use Committee either at GSK or by the
ethical review process at the institution where the work was performed.
26. Dzamko, M.; Deak, M.; Hentati, F.; Reith, A.D.; Prescott, A. R.; Alessi,
D.R.; Nichols, R.J. Biochem. J. 2010, 430, 405.
8. Li, J. Q.; Tan, L.; Yu, J. T. Neurodegener 2014, 9, 47.
9. Dachsel, J. C.; Farrer, M. J. Arch. Neurol.2010, 67, 542.
10. Healy, D. G.; Falchi, M.; O'Sullivan, S. S.; Bonifati, V.; Durr, A.;
Bressman, S.; Brice, A.; Aasly, J.; Zabetian, C. P.; Goldwurm, S.;
Ferreira, J. J.; Tolosa, E.; Kay, D. M.; Klein, C.; Williams, D. R.; Marras,
C.; Lang, A. E.; Wszolek, Z. K.; Berciano, J.; Schapira, A. H.; Lynch, T.;
Bhatia, K. P.; Gasser, T.; Lees, A. J.; Wood, N. W.; International, L. C.
Lancet Neurol. 2008, 7, 583.
11. Bouhouche, A.; Tibar, H.; Ben El Haj, R.; El Bayad, K.; Razine, R.;
Tazrout, S.; Skalli, A.; Bouslam, N.; Elouardi, L.; Benomar, A.;
Yahyaoui, M.; Regragui, W. Parkinsons Dis. 2017, 2017, 2412486.
12. Galatsis, P. Expert. Opin.Ther. Pat. 2017, 27, 667.
13. Christensen, K. V.; Smith, G. P.; Williamson, D. S. Prog. Med. Chem.
2017, 56, 37.
14. Ding, X.; Liu, Q.; Long, K.; Sang, Y.; Stasi, L. P.; Wan, Z.; Xu, Q.; Edge,
C. WO2015113451A1. 2015.
15. Ding, X.; Long, K.; Sang, Y.; Stasi, L. P.; Wan, Z.; Zhao, B.; Edge, C.
WO2015113452A1. 2015.
16. Scott, J. D.; DeMong, D. E.; Greshock, T. J.; Basu, K.; Dai, X.; Harris,
J.; Hruza, A.; Li, S. W.; Lin, S. I.; Liu, H.; Macala, M. K.; Hu, Z.; Mei,
H.; Zhang, H.; Walsh, P.; Poirier, M.; Shi, Z. C.; Xiao, L.; Agnihotri, G.;
Baptista, M. A.; Columbus, J.; Fell, M. J.; Hyde, L. A.; Kuvelkar, R.;
Lin, Y.; Mirescu, C.; Morrow, J. A.; Yin, Z.; Zhang, X.; Zhou, X.;
Chang, R. K.; Embrey, M. W.; Sanders, J. M.; Tiscia, H. E.;Drolet, R. E.;
Kern, J. T.; Sur, S. M.; Renger, J. J.; Bilodeau, M. T.;Kennedy, M. E.;