Small Molecule Inhibits Metal-Dependent and -Independent Multifaceted Toxicity of Alzheimer's Disease

Article abstract of DOI:10.1021/acschemneuro.9b00216

Alzheimer's disease (AD) is one of the most devastating forms of dementia, without reliable treatments to cure, delay the onset, or prevent the disease progression. The proposed toxic mechanisms of AD include amyloidogenesis of amyloid β (Aβ), metal ion dyshomeostasis, redox active metal-Aβ inclusion complex formation, and generation of excessive reactive oxygen and nitrogen species (ROS and RNS). The imbalance in redox homeostasis causes oxidative stress, DNA damage, mitochondrial dysfunction, and inflammation, which collectively become a major hurdle in the development of effective therapeutic agents for multifactorial AD. This necessitates a multifunctional strategy to develop effective therapeutic agents to inhibit multifaceted toxicity. In this context, we report a rational design, synthesis, and detailed study to identify a small molecule multifunctional modulator (MFM) inspired by the human origin tripeptide. The lead, MFM 4, chelates and sequesters metal ions, disrupts their redox cycles, prevents excessive ROS production and oxidative stress, ameliorates oxidative DNA damage and mitochondrial dysfunction, and modulates Nrf2 protein signaling under oxidative stress conditions by eliminating the toxic stress elements. The MFM 4 was found to inhibit metal-dependent and -independent Aβ aggregation and qualified as a suitable candidate to inhibit Aβ-induced neuronal toxicity. The NMR spectroscopy study revealed molecular-level interactions of 4 with Aβ42, which explain the mechanism of aggregation inhibition. Furthermore, 4 effectively inhibited inflammation as revealed by reduction in nitric oxide (NO) production in LPS-activated glial cells. These key features make 4 a potential MFM platform to develop therapeutic agents for metal (Cu, Zn and Fe)-dependent and -independent multifaceted Aβ toxicity of AD.

Full text of DOI:10.1021/acschemneuro.9b00216

Subscriber access provided by UNIV AUTONOMA DE COAHUILA UADEC
Article
Small Molecule Inhibit Metal-Dependent and Independent
Multifaceted Toxicity of Alzheimer’s Disease
Sourav Samanta, Kolla Rajasekhar, Vardhaman Babagond, and Thimmaiah Govindaraju
ACS Chem. Neurosci., Just Accepted Manuscript • DOI: 10.1021/acschemneuro.9b00216 • Publication Date (Web): 29 May 2019
Downloaded from http://pubs.acs.org on May 30, 2019
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 13
ACS Chemical Neuroscience
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 Chemical Neuroscience
Page 2 of 13
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

Page 3 of 13
ACS Chemical Neuroscience
5$./ H< A$. (#/$(/3 A8(1 J$4`P @!++!A/3 5C (#8@+"!#!$B/(8B
/FB/..8%/ ,I (1#!"61 5!(1 &/($+H3/D/'3/'( $'3 &/($+HP
1
2
3
4
5
6
7
8
8'3/D/'3/'( D$(1A$C.) 888? 8'1858( (1/ &/($+H3/D/'3/'(
$'3 &/($+H8'3/D/'3/'( ꢀG $66#/6$(8!' .D/B8/.) 8%? #/3"B/
!F83$(8%/ .(#/.. 8' (1/ '/"#!'$+ B/++.) %? D#!(/B( >JA @#!&
,I ) %8? D#/%/'( &8(!B1!'3#8$+ 3C.@"'B(8!' $'3 !F83$(8%/
3$&$6/) %88? 8'1858( '/"#!6+8$ $B(8%$(8!' $'3 D#!%83/ $'(8H
8'@+$&&$(!#C /@@/B(.) $'3 %888? D#!%83/ !%/#$++
'/"#!D#!(/B(8!' $'3 #/.B"/ '/"#!'$+ B/++. @#!& &/($+H
$B83 (! !5($8' (1/ ($#6/( B!&D!"'3 HE 71/ QH>!D$ &/(1C+
/.(/# A$. (#/$(/3 A8(1 74>L S+) $'3 (1/ #/."+(8'6 D#!3"B(
A$. B!"D+/3 A8(1 M&!BH`8.=7#(?HI` ".8'6 `47a $'3
`I4( 8' >LM @!++!A/3 5C M&!BH3/D#!(/B(8!') A18B1 6$%/
(1/ 8'(/#&/38$(/ G<E 71/ 8'(/#&/38$(/ G< A$. (#/$(/3 A8(1
4!BH9+CHI` 8' >LM @!++!A/3 5C (#/$(&/'( A8(1
e
(#8@+"!#!$B/(8B $B83 $'3 $&&!'8"& @+"!#83/ =J`P MH? (!
3/D/'3/'( $'3
&"+(8@$B/(/3 (!F8B8(CE
&/($+H8'3/D/'3/'(
ꢀGH&/38$(/3
!5($8' (1/ ($#6/( B!&D!"'3 GE 71/ 8'(/#&/38$(/ G< A$.
(#/$(/3 A8(1 .$+8BC+$+3/1C3/ 8' >LM (! !5($8' B18@@ 5$./
J&<KE 71/ B18@@5$./ 8'(/#&/38$(/ &< A$. (#/$(/3 A8(1
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
e
J$4`P) @!++!A/3 5C (#8@+"!#!$B/(8B $B83 $'3 J`P MH (!
#(!/8"! A., ,-!$/!!-+.
!5($8' (1/ ($#6/( B!&D!"'3E 71/ 8'(/6#8(C !@ $++ (1/
8'(/#&/38$(/. $'3 (1/ @8'$+ B!&D!"'3. =6H&? A$.
B!'@8#&/3 5C `861 R/#@!#&$'B/ Q8N"83 S1#!&$(!6#$D1C
=`RQS?) JL, $'3 1861H#/.!+"(8!' &$.. .D/B(#!&/(#C
=`,L ?E
,5912: !;4<;52= >? .<;34<@ "41A5A;1B5C1:9A145B !D<@@
E>@5F3@5CE0E7 _/ 3/.86'/3 $ ./( !@ .&$++ &!+/B"+/. (!
83/'(8@C
$ D!(/'(8$+ LML 5C "'3/#($08'6 .(#$(/68B
.(#"B("#$+ $'3 @"'B(8!'$+ &!38@8B$(8!'. (! 9`*) $ '$("#$+
(#8D/D(83/ 0'!A' @!# "&D(//' '"&5/# !@ 58!+!68B$+
@"'B(8!'. 8' 1"&$'.E[W)[Z I'/ !@ (1/ &$-!# #/%/+$(8!'. 1/#/
8. (1$( fOWW J6gQ !@ 9`* 8. @!"'3 8' $3"+( 1"&$' ./#"&)
A18B1 3/B#/$./. (! hXW J6gQ A8(1 $68'6E[W ,/&$#0$5+C)
9`* /F1858(. 1861/# 58'38'6 $@@8'8(C =*$ ZWZP? B!&D$#/3
(! ꢀKPO =*$ ZW]? @!# S"KK) 5"( +!A/# (1$' (1$( !@
&/($++!D#!(/8'. =*$ ZWZ\ H ZWZV? 8' (1/ 58!+!68B$+ &8+8/"E[Z K'
!(1/# A!#3.) 9`* B$' /@@/B(8%/+C ./N"/.(/# S"KK @#!& ꢀKPOH
S"KK B!&D+/F A8(1!"( 8'(/#@/#8'6 A8(1 B!DD/#H5$./3
&/($++!CD#!(/8'.) $ 1861+C 3/.8#$5+/ D#!D/#(C (1$( 1$. 5//'
/FD+!8(/3 8' !"# /$#+8/# A!#0E[W `!A/%/#) 9`* $'3 8(.
B!'-"6$(/. @"'B(8!' $. $'(8HA> $6/'(. !'+C 8' (1/ D#/./'B/
!@ S"KK $'3 $#/ 6#!..+C 8'/@@/B(8%/ 8' (1/ 8'1858(8!' !@ ,I
6/'/#$(8!') !F83$(8%/ .(#/.. $'3 '/"#!8'@+$&&$(8!' $#8./
@#!& M/H3/D/'3/'( $'3 &/($+ =S"KK $'3 M/KKK?H8'3/D/'3/'(
D#!B/../.E K' (18. B!'(/F() A/ /&5$#0/3 !' D/#(8'/'( $'3
"'8N"/ .(#"B("#$+ $'3 @"'B(8!'$+ &!38@8B$(8!'. (! 9`* (!
6/'/#$(/ '!%/+ .&$++ &!+/B"+/HLML. B$D$5+/ !@
&!3"+$(8'6 (1/ &/($+H3/D/'3/'( $'3 8'3/D/'3/'(
6/'/#$(8!' !@ /FB/..8%/ ,I $. A/++ $. ꢀK $66#/6$(8!') $'3
$I5@<;1>: >? #5B>LCAF;1M5 E5;<@ ->:97 S!!#38'$(8!' !@
#/3!FH$B(8%/ &/($+ 8!'. =S"KK $'3 M/KKK? A8(1 ꢀK 1$. 5//'
.1!A' (! /'1$'B/ (1/ $66#/6$(8!' $'3 .($58+8;$(8!' !@ (1/
!+86!&/#8B .($(/ $'3 6/'/#$(8!' !@ /FB/..8%/ ,I (1#!"61
$ B!'(8'"!". #/3!F BCB+8'6 D#!B/..E])[OH[P 71/ /@@/B(8%/
B1/+$(8!' $'3 ./N"/.(#$(8!' !@ #/3!FH&/($+ 8!'. @#!& ꢀKH
&/($+ 8'B+".8!' B!&D+/F $'3 ."5./N"/'( #/3"B(8!' !@
/FB/..8%/ ,I $#/ (1/ 0/C .(#$(/68/. (! $&/+8!#$(/ (1/
NKO GVP
NAO
GVP
PVT
PVR
PVL
PV8
PVP
GVP
PVT
PVR
PVL
PV8
PVP
WHX
WHX
D
F
E
<
D
F
E
<
PVT
PVR
PVL
PV8
PVP
N%*'&*.8".,%* OQ$P
N%*'&*.8".,%* OQ$P
N[O PVP
N"O PVR
R
B!'(#!++8'6
(1/
#/+$(/3
!F83$(8%/
.(#/..
$'3
PVP
DPVQ
DGVP
DGVQ
D8VP
D8VQ
DMVP
DPV8
'/"#!8'@+$&&$(8!') (1/#/5C D#!(/B(8'6 >JA $'3
&8(!B1!'3#8$E K' 9`*) 6+CB8'/ =9? $'3 18.(838'/ =`? $#/
8'38.D/'.$5+/ @!# S"KK B1/+$(8!' A18+/ +C.8'/ =*? &!.(+C
$..8.(. &/&5#$'/ $'B1!#8'6 (! (#$'.D!#( S"KK /@@8B8/'(+C
8'.83/ (1/ B/++.E[Z K' !"# 3/.86') (1/ &/($+ 8!' B1/+$(8!'
D#!D/#(C !@ 9` =8' 9`*? A$. 8'(/6#$(/3 A8(1 D!+CD1/'!+8B
&!8/(8/. ."B1 $. QH>IRA $'3 3!D$&8'/ (! !5($8'
B!&D!"'3. 6 $'3 G = /(HZ) M86"#/ Z?E 7! @"#(1/# /'1$'B/ (1/
&/($+ B1/+$(8!' $58+8(C (!A$#3. S" $'3 M/) $'(8!F83$'(
D#!D/#(C $'3 8'1858(8!' !@ ꢀK $66#/6$(8!') 6+CB8'/ A$.
."5.(8("(/3 A8(1 $ .$+8BC+$+3/1C3/ &!8/(C (! D#!3"B/ H $'3
& = /(HO) M86"#/ Z?E
L
PVL
DPVL
DPVR
DPVT
DGVP
P
8
L
R
T
P
V
G
V
V
V
V
V
P
P
P
P
P
+9%/ :%*)+
8VP
GVQ
GVP
PVQ
PVP
8
PV8
P
PVP
P
Q
P
Q
P
V
8
V
V
V
V
P
P
G
G
$%#"8 ;".,%
4,!& O!,*P
4,!& O!,*P
N!O
N\O
S
GVP
]$^[3_
]$^!(_
]$_
]$_
R
PVT
PVR
]$^8[3_
]$^8!(_
Q
L
]$^M[3_
]$^M!(_
M
8
G
P
PVL
PV8
PVP
!=:;I5919 >? $>DA>3:B9 6H&7 71/ .C'(1/(8B #!"(/
@!++!A/3 @!# (1/ D#/D$#$(8!' !@ 6H& 8. .1!A' 8' M86"#/ Z4E
M&!BH`8.=7#(?HI` A$. B!"D+/3 (! 3!D$&8'/ ".8'6 `47a
$'3 `I4( 8' >LM) $'3 (1/ D#!3"B( A$. ."5-/B(/3 (!
!A7
!A7
5"#3/' 012345 G7 S1/+$(8!' !@ #/3!F $B(8%/ &/($+ 8!'. =S"KK
$'3 M/KKK? 5C B!&D!"'3. 6H&E A? 71/ D+!( !@ $5.!#5$'B/
8'(/'.8(C $( \]\ '& $'3 ^VW '& @!# S"KK B!&D+/F !@ 6 H& $'3
9`*E 4? 71/ D+!( !@ $5.!#5$'B/ 8'(/'.8(C $( \WW '& @!# M/KKK
B!&D+/F !@ B!&D!"'3. 6H& $'3 9`*E S? $'3 >? K7S 58'38'6
8.!(1/#&. !@ & S"KK $'3 &HM/KKK B!&D+/F/.) #/.D/B(8%/+CE b? $'3
M&!BH3/D#!(/B(8!'
(!
!5($8'
18.(838'/H3!D$&8'/
B!'-"6$(/ 6<E 71/ 8'(/#&/38$(/ 6< A$. B!"D+/3 (! 4!BH9+CH
I` 8' >LM @!++!A/3 5C 4!B) $'3 7#(H3/D#!(/B(8!' 6$%/
B!&D!"'3 6E J/F() 8'(/#&/38$(/ 6< A$. B!'-"6$(/3 A8(1
.$+8BC+$+3/1C3/ 8' >LM (! !5($8' B18@@ 5$./ H<E 71/ B18@@
"
ACS Paragon Plus Environment

ACS Chemical Neuroscience
Page 4 of 13
M? LAQ>K &$.. $'$+C.8. !@ i&HS"KKj $'3 i&HM/KKKj B!&D+/F/.)
#/.D/B(8%/+CE
S"KK $'3 M/KKK (! ($B0+/ (1/ &/($+H3/D/'3/'( ,I
1
2
3
4
5
6
7
8
6/'/#$(8!' $'3 #/+$(/3 $3%/#./ /@@/B(.E 71".) A/ /FD+!#/3
(1/ /@@/B( !@ !"# B!&D!"'3. !' (1/ #/3!FH&/($+ 3/D/'3/'(
ꢀK (!F8B8(C =,I 6/'/#$(8!' $'3 !F83$(8%/ .(#/..?E
!@ &"+(8@$B/(/3 ꢀK (!F8B8(C 8'B+"38'6 !F83$(8%/ .(#/..E[[H[^
71/ B1/+$(8'6 $58+8(C !@ (1/ B!&D!"'3. 6H& (!A$#3. #/3!FH
$B(8%/ &/($+ 8!'. =S"KK $'3 M/KKK? A$. .("38/3 5C (1/
$5.!#D(8!' &/$."#/&/'(.E S!&D!"'3. 6 $'3 G A8(1 9`
E5;<@CB5A5:B5:; A:;1>L1B<:; A99<=7 71/ 8'B+".8!'
B!&D+/F !@ #/3!FH$B(8%/ &/($+ 8!'. =S"KK $'3 M/KKK? 8' ꢀK
.D/B8/. 8'.(86$(/. (1/ M/'(!' (CD/ #/$B(8!' (! 6/'/#$(/
/FB/..8%/ ,I =`OIO $'3 I`m? $'3 +/$38'6 (! !F83$(8%/
.(#/.. $'3 #/+$(/3 (!F8B8(CE])Z^)[\ 71/#/@!#/) B1/+$(8!' !@
#/3!FH$B(8%/ &/($+ 8!'. =S"KK $'3 M/KKK? $'3 0//D8'6 (1/&
8' (1/ #/3!FH3!#&$'( .($(/ "'3/# #/3"B8'6 /'%8#!'&/'( 8.
B#"B8$+ (! D#/%/'( /FB/..8%/ ,I 6/'/#$(8!' $'3 !F83$(8%/
.(#/..E\)OW)[])PW _/ D/#@!#&/3 !& :!-#" $'3 !& '*6676"
$'(8!F83$'( $..$C. ".8'6 #/3!F &/($+ 8!' =S"KK !# M/KKK? $'3
$.B!#5$(/ =A.B? (! %$+83$(/ (1/ #/3!FH.8+/'B8'6 $58+8(C !@
!"# B!&D!"'3. =6H&?E M86"#/ [A .1!A. (1$( .$&D+/
8'B"5$(/3 A8(1 S"KK 8' (1/ $5./'B/ !@ !"# B!&D!"'3.
.1!A/3 &$F8&"& [HSSA @+"!#/.B/'B/ /&8..8!' =ZWWY?E
A338(8!' !@ B!&D!"'3. 6H& =!OW ML? B!'.83/#$5+C #/3"B/3
(1/ @+"!#/.B/'B/ /&8..8!' (! hZWY) A1/#/$. (1/ B!'(#!+
9`* .1!A/3 .86'8@8B$'( [HSSA @+"!#/.B/'B/ =VZY? $( $
B!'B/'(#$(8!' $. 1861 $. \W MLE 7! B1/B0 (1/ D#!3"B(8!'
!@ /FB/..8%/ I`m &!#/ /@@8B8/'(+C $. B!&D$#/3 $( +!A/#
B!'B/'(#$(8!' =ZW JL? !@ B!&D!"'3. H $'3 &) (1/ .$&D+/.
.1!A/3 [HSSA @+"!#/.B/'B/ /&8..8!' !@ PX $'3 \PY)
#/.D/B(8%/+C B!&D$#/3 (! X[ $'3 VXY @!# 6 $'3 G)
#/.D/B(8%/+C) A18B1 8'38B$(/. (1$( H $'3 & A/#/ (1/ ."D/#8!#
(! !(1/#.) A18B1 .1!A/3 &!3/#$(/ $B(8%8(C =M86"#/ ^?E
J/F() A/ $../../3 (1/ /@@/B( !@ B!&D!"'3. 6H& !' (1/
D#!3"B(8!' !@ I`m @#!& (1/ M/'(!' (CD/ #/$B(8!' !@ M/KKK
=M86"#/ V?E A. /FD/B(/3) (1/ 38.D#!D!#(8!'$(8!' #/$B(8!'
!BB"##/3 8' (1/ .$&D+/ B!'($8'8'6 `OIO $+!'/ $'3 383 '!(
.1!A $'C [HSSA @+"!#/.B/'B/ /'1$'B/&/'(E `!A/%/#) 8'
(1/ D#/./'B/ !@ M/KKK) $ .(#!'6 @+"!#/.B/'B/ /'1$'B/&/'(
A$. !5./#%/3 !A8'6 (! (1/ D#!3"B(8!' !@ /FB/.. I`m
=M/'(!' (CD/ #/$B(8!'?E ,/&$#0$5+C) .$&D+/. (#/$(/3 A8(1
B!&D!"'3. H $'3 & .1!A/3 \W $'3 \\Y #/3"B(8!')
#/.D/B(8%/+C) 8' [HSSA @+"!#/.B/'B/ /&8..8!'c 6) G $'3 9`*
=Z^) Z\ $'3 Z[Y #/3"B(8!') #/.D/B(8%/+C? .1!A/3 &8'8&$+
#/3"B(8!' B!&D$#/3 (! (1/ B!'(#!+ =ZWWY?E 718. 8. $ B+/$#
8'38B$(8!' (1$( H $'3 &H5!"'3 M/KKK A$. '!( 8'%!+%/3 8' (1/
#/3!F D#!B/.. (! 6/'/#$(/ I`m 8' (1/ D#/./'B/ !@ `OIOE I'
(1/ !(1/# 1$'3) B!&D!"'3. 6) G $'3 9`* A/#/ @!"'3 (!
1$%/ &8'8&$+ 8'(/#@/#/'B/ 8' (1/ M/KKK #/3!F D#!B/.. @!#
B1/B08'6 ,I 6/'/#$(8!' =M86"#/ V?E
38D/D(83/ "'8( /F1858(/3 5#!$3 $5.!#D(8!' 5$'3. =k&$F
l
\]\ '&? 8' (1/ D#/./'B/ !@ S"KK) A18B1 8'38B$(/3 (1/
@!#&$(8!' !@ B!##/.D!'38'6 38.(!#(/3 .N"$#/HD+$'$#
B!&D+/F/. +80/ 9`* =M86"#/ OA?E[V I' (1/ !(1/# 1$'3) H
$'3 & .1!A/3 (A! 38.(8'B( B1$#$B(/#8.(8B $5.!#D(8!' 5$'3.
A8(1 (1/ &$F8&$ $( PZ\ '& $'3 ^]W '&) #/.D/B(8%/+CE 71/
.(#!'6 $5.!#D(8!' 8'(/'.8(C $( PZ\ '& $'3 $ +$#6/
5$(1!B1#!&8B .18@( =fV\ '&? B!&D$#/3 (! 9`* =k&$F l \]\
'&? 8'38B$(/3 (1/ D!..85+/ 8'%!+%/&/'( !@ D1/'!+8B
1C3#!FC+ 6#!"D. 8' S"KK B1/+$(8!' = "DD!#(8'6 K'@!#&$(8!')
M86"#/ Z?E b%83/'(+C) Z` JL, .D/B(#$ .1!A/3 (1/
3/D#!(!'$(8!' !@ $&83/ 1C3#!6/' $( XE[\ DD&) A18B1
B!'@8#&/3 (1/ 8'%!+%/&/'( !@ 3/D#!(!'$(/3 $&83/
'8(#!6/' !@ & 8' B!&D+/F$(8!' A8(1 S"KK) .8&8+$# (! 9`* e
S"KK B!&D+/F$(8!' =M86"#/ O?EPO J/F() A/ $../../3 (1/
$58+8(C !@ !"# B!&D!"'3. (! B1/+$(/ M/KKK $'3 8'1858( 8(.
#/3!F $B(8%8(C) $' 8'1/#/'( 3#$A5$B0 !@ 9`*E ,/&$#0$5+C)
$++ (1/ B!&D!"'3. =6H&? .1!A/3 8'(/'./ 5#!$3 $5.!#D(8!'
5$'3. 8' (1/ %8.85+/ #/68!' =k&$F l \WW '&? 8' (1/ D#/./'B/
!@ M/KKK =M86"#/ O4?E 71/ '/A $5.!#D(8!' 5$'3 8' (1/ %8.85+/
#/68!' A$. $((#85"(/3 (! (1/ !#&$(8!' !@ M/KKK B!&D+/F)
D!..85+C (1#!"61 1C3#!FC+ 6#!"D.E[X ,/&$#0$5+C) 9`* 383
'!( .1!A $'C $5.!#5$'B/ B1$'6/ 8' (1/ D#/./'B/ !@ M/KKK
=M86"#/ [?E K' 8.!(1/#&$+ (8(#$(8!' B$+!#8&/(#C =K7S?
&/$."#/&/'( $++ (1/ B!&D!"'3. 8'B+"38'6 9`* .1!A/3
/'3!(1/#&8B 58'38'6 A8(1 S"KK =M86"#/ OS? $'3
38..!B8$(8!' B!'.($'(. =*3? 8' (1/ '$'!&!+$# #$'6/ :!LE)
[[EP] F ZWH]) PPEO\ F ZWH]) O^EVW F ZWH]) OOEZZ F ZWH] $'3 Z]EPP F
ZWH] L @!# 9`* $'3 B!&D!"'3. 6H&) #/.D/B(8%/+C =M86"#/
P?E I' (1/ !(1/# 1$'3) /F!(1/#&8B 58'38'6 8'(/#$B(8!'
A$. !5./#%/3 @!# (1/ B!&D+/F$(8!' !@ M/KKK =M86"#/ O>? A8(1
B!&D!"'3. 6H& A8(1 38..!B8$(8!' B!'.($'(. =*3? 8' (1/
'$'!&!+$# #$'6/ :!LE) [PE[W F ZWH]) OVE^W F ZWH]) \EW\ F ZWH]
$'3 ^E^^ F ZWH] L) #/.D/B(8%/+C =M86"#/ \?E "#D#8.8'6+C)
9`* $+.! .1!A/3 58'38'6 8'(/#$B(8!' A8(1 M/KKK 8' (1/ K7S
&/$."#/&/'(. A8(1 (1/ 38..!B8$(8!' B!'.($'( %$+"/ !@ ZE[V
F ZWH] LE K' .D8(/ !@ (1/ 58'38'6 8'(/#$B(8!' !5./#%/3 8' K7S
3$($) 9`* A$. @!"'3 (! 5/ 8'/@@/B(8%/ 8' &!3"+$(8'6 (1/
6/'/#$(8!' !@ ,I (1#!"61 M/'(!' (CD/ #/$B(8!') D!..85+C
3"/ (! 8(. 8'$58+8(C (! 0//D M/KKK 8' $ #/3!FH3!#&$'( .($(/
=M86"#/ V) ZW?E M"#(1/#) &$.. .D/B(#!&/(#C 3$($
."DD!#(/3 (1/ B!&D+/F @!#&$(8!' 5/(A//' B!&D!"'3. 6H&
$'3 (1/ &/($+ 8!' =S"KK $'3 M/KKK? =7$5+/ Z?E LAQ>K
$'$+C.8. !@ B!&D!"'3 & $'3 S"KK .1!A/3 &g; D/$0.
B!##/.D!'38'6 (! .(#!'6 iLeS"KKj) iLeOS"KKj $'3
iLe[S"KKj 8'(/#$B(8!'. =\ZVEZZ) \XWEWP $'3 ^POE]V)
#/.D/B(8%/+C? =M86"#/ Ob?E 8&8+$#+C) B!&D+/F$(8!' !@ & $'3
M/KKK A$. B!'@8#&/3 5C (1/ &g; D/$0. B!##/.D!'38'6 (!
iLeM/KKKj) iLeOM/KKKj $'3 iLe[M/KKKj 8'(/#$B(8!'. =\ZWEZO)
\^^EW\ $'3 ^OZE]]) #/.D/B(8%/+C? =M86"#/ OM?E 71/
$5.!#5$'B/) JL,) K7S $'3 &$.. $'$+C.8. 3$($ (!6/(1/#
"'3/#.B!#/ (1/ @$B( (1$( A/ $B18/%/3 (1/ @8#.( 6!$+ !@
3/.86'8'6 B!&D!"'3. (1$( B$' B1/+$(/ 5!(1 #/3!FH$B(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
>JA !F83$(8%/ 3$&$6/ 5C /+/%$(/3 ,I "'3/# A>
B!'38(8!' 8. !'/ !@ (1/ &!.( 3#/$3@"+ B!'./N"/'B/. (1$(
$66#$%$(/ (1/ '/"#!'$+ (!F8B8(CE[W 71/ B1/&8B$+ #/$B(8!' !@
,I A8(1 >JA B$"./3 5#/$08'6 !@ (1/ D1!.D1$(/
5$B05!'/ !# '"B+/!5$./ &!38@8B$(8!'.) +/$38'6 (! B/++"+$#
3/$(1E\ _/ $../../3 (1/ $58+8(C !@ B!&D!"'3. 6H& $'3 9`*
(! D#!(/B( (1/ >JA @#!& !F83$(8%/ 3$&$6/ ".8'6 D+$.&83
>JA =DaSZ]? $. $ &!3/+ .C.(/& =M86"#/ [4?E A6$#!./ 6/+
3$($ .1!A/3 (1$( >JA .$&D+/ (#/$(/3 A8(1 S"KKHA.B =B(#+?
/F1858(/3 fZWWY '!'HB!8+/3 =JS? @!#& A18+/ D>JA =R4
(#/$(/3? B!'($8'/3 fZPY JS =/F8.(8'6 &!.(+C $.
."D/#B!8+/3 @!#&) S?) A18B1 $((#85"(/3 (! !F83$(8%/ >JA
3$&$6/ 5C (1/ !& ,!-7 6/'/#$(/3 I`mE a'3/# .8&8+$#
#
ACS Paragon Plus Environment

Page 5 of 13
ACS Chemical Neuroscience
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 Chemical Neuroscience
Page 6 of 13
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

Page 7 of 13
ACS Chemical Neuroscience
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 Chemical Neuroscience
Page 8 of 13
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

Page 9 of 13
ACS Chemical Neuroscience
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 Chemical Neuroscience
Page 10 of 13
$..$CEZ] 71/ B/++. 8'B"5$(/3 A8(1 `OIO /F1858(/3 OPY
8'@+$&&$(!#C D#!D/#(8/. $'3 8'1858(8!' !@ &/($+ 8!'H
1
2
3
4
5
6
7
8
,1!ZO[ @+"!#/.B/'B/ /&8..8!' $. B!&D$#/3 (! (1/
"'(#/$(/3 B!'(#!+ =ZWWY?) A18B1 8. 8'38B$(8%/ !@ (1/
!F83$(8%/ .(#/..H8'3"B/3 &8(!B1!'3#8$+ 3C.@"'B(8!' =LLR
#/3"B(8!'?E 71/ B/++. (#/$(/3 A8(1 `OIO $'3 B!&D!"'3 &
=ZW) OW $'3 \W ML? .1!A/3 /'1$'B/3 ,1!ZO[ @+"!#/.B/'B/
/&8..8!' =8&D+C8'6 (1/ B!##/.D!'38'6 8&D#!%/&/'( 8'
LLR? "D (! O]) P\ $'3 ^WY #/.D/B(8%/+C) 5C .B$%/'68'6 (1/
(!F8B #$38B$+.) A18+/ 9`* =\W ML? @$8+/3 (! .1!A $'C
.86'8@8B$'( 8&D#!%/&/'( 8' (1/ LLR =M86"#/ ^4) S?E 71/./
!5./#%/3 #/."+(. ."66/.( (1$( & #/.B"/3 &8(!B1!'3#8$ 5C
#/.(!#8'6 8(. LLR) A18B1 A$. $((#85"(/3 (! /@@8B8/'(
8'1858(8!' !@ !F83$(8%/ .(#/.. 5C .B$%/'68'6 (1/ /FB/.. ,I
D#!3"B/3 8' (1/ B/++. =M86"#/ V?E
3/D/'3/'( $'3 8'3/D/'3/'( AKPO $66#/6$(8!' &$0/
B!&D!"'3 & $ 1861+C 3/.8#$5+/ LML @!# 3/%/+!D8'6
(1/#$D/"(8B $6/'(. (! $&/+8!#$(/ &"+(8@$B/(/3 AK (!F8B8(C
8' A> =M86"#/ V?E _/ 1$%/ A8('/../3 ./#8/. !@ 3#"6
B$'383$(/. @$8+/3 $( %$#8!". .($6/. !@ B+8'8B$+ (#8$+. D!..85+C
3"/ (! (1/8# 8'$58+8(C (! 8'(/#@/#/ A8(1 &"+(8@$B/(/3 (!F8B8(C
!@ A>E K' (18. B!'(/F() !"# &"+(8@"'B(8!'$+ &!3"+$(!#
=LML? .(#$(/6C 8. $'(8B8D$(/3 (! 8'.D8#/ (1/ 3/%/+!D&/'(
!@ D!(/'(8$+ (1/#$D/"(8B B$'383$(/. (! (#/$( A>) 8' (1/ '/$#
@"("#/E
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
E("S+,!
K' B!'B+".8!') A/ 1$%/ 3/&!'.(#$(/3 !"# #$(8!'$+ 3/.86'
$'3 .C'(1/.8. !@ '$("#$+ (#8D/D(83/H8'.D8#/3 .&$++
!=:;I5919 >? $>DA>3:B9 6C&7 7! $ .(8##/3 .!+"(8!' !@
M&!BH`8.=7#(?HI` =[EW 6) PEX &&!+? 8' >LM =^ &+? $( W
dS) >KRbA =ZEW[ &Q) \EX &&!+?) `47a =OEOW6) \EX
&&!+?$'3 `I4( =WEVX 6) \EX &&!+? A/#/ $33/3E 71/
#/$B(8!' &8F("#/ A$. 0/D( @!# .(8##8'6 $5!"( Z\ &8' "'3/#
'8(#!6/' $(&!.D1/#/E 71/ [)PH381C3#!FCD1/'/(1C+$&8'/
=3!D$&8'/? =ZEX[ 6) ]E^X &&!+? $'3 &/(1C+ [H=[)PH58.=(/#(H
5"(C+38&/(1C+.8+C+!FC?D1/'C+?HOH$&8'!D#!D$'!$(/ =[EW 6)
^EX &&!+? A/#/ $33/3 (! $5!%/ .!+"(8!') $'3 (1/ #/$B(8!'
A$. +/@( (! .(8# @!# \H^ 1E A@(/# (1/ B!&D+/(8!' !@ (1/
#/$B(8!' =&!'8(!#/3 5C 7QS?) (1/ .!+%/'( A$. #/&!%/3)
$'3 (1/ B#"3/ A$. 38+"(/3 A8(1 A$(/# =[W &Q?) $'3 (1/
#/.83"/ A$. /F(#$B(/3 8'(! b(IAB =[ t [W &Q?E 71/
B!&58'/3 !#6$'8B D1$./ A$. A$.1/3 A8(1 A$(/# =Z t [W
&Q? $'3 5#8'/ =Z t [W &Q?E71/ !#6$'8B +$C/#. A/#/
B!++/B(/3) 3#8/3 !%/# $'1C3#!". J$O IP $'3 /%$D!#$(/3 8'
%$B""& (! $@@!#3 (1/ B#"3/ D/D(83/E 71/ D#!3"B(. A$#/
D"#8@8/3 5C B!+"&' B1#!&$(!6#$D1C ".8'6 >SL $'3
&/(1$'!+ $. /+"/'(E J/F() 6< =WE\ 6) WE][ &&!+? $'3 GN =ZE\
6) ZEX[ &&!+? A$. 38..!+%/3 8' >LM =[ &+?) A18B1 A$.
B!!+/3 (! W dS @!++!A/3 5C >KRbA =WE[[ &Q) ZEXV &&!+? )
`47a =WEPOV 6) ZEZO &&!+? $'3 `I4( =WEZV 6) ZEZO &&!+?
A/#/ $33/3E 71/ #/$B(8!' &8F("#/ A$. 0/D( @!# .(8##8'6
$5!"( Z\ &8' "'3/# '8(#!6/' $(&!.D1/#/ $'3 4!BH9+CHI`
=WEO 6) ZEZO &&!+? A$. $33/3 (! $5!%/ .!+"(8!' $'3 (1/
#/$B(8!' A$. +/@( (! .(8# @!# \H^ 1 $( #!!& (/&D/#$("#/E A@(/#
(1/ B!&D+/(8!' !@ (1/ #/$B(8!' =&!'8(!#/3 5C 7QS?)
.!+%/'( A$. #/&!%/3 $'3 (1/ B#"3/ A$. 38+"(/3 A8(1 A$(/#
=O\ &Q? $'3 #/.83"/ A$. /F(#$B(/3 8'(! b(IAB =[t O\
&Q?E71/ B!&58'/3 !#6$'8B D1$./ A$. A$.1/3 A8(1 A$(/#
=Z t [W &Q? $'3 5#8'/ =Z t O\ &Q?E 71/ !#6$'8B +$C/#. A/#/
B!++/B(/3) 3#8/3 !%/# $'1C3#!". J$O IP $'3 /%$D!#$(/3E
718. A$. D"#8@8/3 5C B!+"&' B1#!&$(!6#$D1C ".8'6
38B1+!#!&/(1$'/ =>SL? $'3 &/(1$'!+ $. /+"/'(E M8'$++C)
(1/ $5!%/ B#"3/ B!&D!"'3. 6HG A/#/ D"#8@8/3 ".8'6 $
#/%/#./HD1$./ =,R? ./&8HD#/D$#$(8%/ `RQS !' SZX B!+"&'
$( PW dSE M"#(1/#) (! .C'(1/.8. B!&D!"'3. HH&
8'(/#&/38$(/. 6N =WEP 6) WEV\ &&!+? $'3 GN =ZEW 6) ZEOO
&&!+? 8' $B/(!'8(#8+/ =ZW &Q? A/#/ $33/3 OH
1C3#!FC5/';$+3/1C3/ =WEO &Q) ZEX[ &&!+? >KRbA =WE\ &Q)
OEPP &&!+?) 71/ #/$B(8!' &8F("#/ A$. 0/D( @!# .(8##8'6
$5!"( ZW &8' $( #!!& (/&D/#$("#/ "'3/# '8(#!6/'
$(&!.D1/#/ $'3 (18. #/$B(8!' &8F("#/ A$. 1/$(/3 "D (! ^\
dS $'3 (1/ #/$B(8!' A$. +/@( (! .(8# @!# ZO 1E A@(/# (1/
B!&D+/(8!' !@ (1/ #/$B(8!' =&!'8(!#/3 5C 7QS?) (1/
&!+/B"+/HLML.
5C
."BB/..@"++C
8'(/6#$(8'6
&"+(8@"'B(8!'$+ D#!D/#(8/. (! ($#6/( &/($+H3/D/'3/'( $'3
8'3/D/'3/'( &"+(8@$B/(/3 ꢀK (!F8B8(C $..!B8$(/3 A8(1
A+;1/8&/#<. 38./$./ D$(1!+!6CE 71/ 3/($8+/3 /%$+"$(8!'
.("3C #/%/$+/3 (1$( B!&D!"'3 & A$. (1/ &!.( D!(/'( $'3
/@@/B(8%/ &!3"+$(!# !@ &"+(8@$B/(/3 ꢀK (!F8B8(C
/'B!&D$..8'6 &/($+ 8!' 3C.1!&/!.($.8.) &/($+H
3/D/'3/'( ,I 6/'/#$(8!') &/($+H3/D/'3/'( $'3
8'3/D/'3/'( AKPO (!F8B8(C) !F83$(8%/ .(#/..) >JA 3$&$6/)
&8(!B1!'3#8$+ 3C.@"'B(8!' $'3 '/"#!8'@+$&&$(8!') A18B1
$#/ (1/ &$-!# 1"#3+/. 8' (1/ 3/%/+!D&/'( !@ (1/#$D/"(8B
$6/'(. @!# &"+(8@$B(!#8$+ A>E 71/ $5.!#D(8!' .("38/.) K7S
&/$."#/&/'(.) $'3 &/($+H3/D/'3/'( $'(8!F83$'( $..$C.
3/&!'.(#$(/3 (1$( & B1/+$(/3 #/3!FH$B(8%/ &/($+ 8!'. $'3
0/D( (1/& 8' (1/ #/3!FH3!#&$'( .($(/ (! $##/.( (1/
D#!3"B(8!' !@ /FB/..8%/ ,I ) (1/#/5C &!3"+$(8'6
!F83$(8%/ .(#/..E K'(/#/.(8'6+C) (1/ '$'!&!+$# $@@8'8(8/. !@
LML (!A$#3. #/3!F &/($+ 8!'. =S"KK $'3 M/KKK?) $. #/%/$+/3
5C (1/ K7S 3$($) B!'@8#&/3 (1/ ./N"/.(#$(8!' !@ (1/./ &/($+
8!'.E 71/ D!+CD1/'!+8B &!8/(C !@ & B!'(#85"(/3 (! /@@8B8/'(
#$38B$+ N"/'B18'6 $58+8(C $. .1!A' 5C 7#!+!F) A47 $'3
>RR` $..$C.E M"#(1/#) 717 @+"!#/.B/'B/ $..$C) 7bL $'3
3!( 5+!( $'$+C.8. B+/$#+C #/%/$+/3 (1$( LML /@@8B8/'(+C
8'1858(/3 5!(1 &/($+H3/D/'3/'( $'3 8'3/D/'3/'( AKPO
$66#/6$(8!' 8' 5!(1 8'1858(8!' $'3 38..!+"(8!' $..$C.E 71/
JL, .("3C #/%/$+/3 &!+/B"+$#H+/%/+ 8'(/#$B(8!'. 5/(A//'
(1/ LML $'3 AKPO (1#!"61 1C3#!6/' 5!'38'6 $'3
1C3#!D1!58B 8'(/#$B(8!'.) A18B1 38.#"D( (1/ $../&5+C !@
AKPO (! @!#& (!F8B $66#/6$(/.E M"#(1/#) !& '*6676" .("38/.
A/#/ 8' 6!!3 $6#//&/'( $'3 ."DD!#(/3 (1/ !& :!-#" $..$C.)
A1/#/ LML #/.B"/3 B/++. @#!& &"+(8@$B/(/3 AK (!F8B8(C 5C
&!3"+$(8'6 (1/ &/($+H3/D/'3/'( $'3 8'3/D/'3/'( AKPO
$66#/6$(8!') ,I ) !F83$(8%/ .(#/.. $'3 >JA 3$&$6/E
,/&$#0$5+C) & #/.B"/3 &8(!B1!'3#8$ @#!& 3C.@"'B(8!' 5C
#/.(!#8'6 8(. LLR) A18B1 A$. $((#85"(/3 (! /@@8B8/'(
.B$%/'68'6 !@ /FB/..8%/ ,I $'3 8'1858(8!' !@ !F83$(8%/
.(#/.. 8' (1/ B/++.) @"#(1/# ."DD!#(/3 5C (1/ #/.(!#$(8!' !@
'"B+/$# $'3 BC(!.!+8B J#@O +!B$+8;$(8!' "'3/# (1/ !F83$(8%/
.(#/.. B!'38(8!' =M86"#/ V?E K' $338(8!') LML .86'8@8B$'(+C
#/3"B/3 (1/ QR H&/38$(/3 6+8$ B/++. $B(8%$(8!') JI
D#!3"B(8!') $'3 8'@+$&&$(8!'E I%/#$++) (1/ 6!!3 B/++
%8$58+8(C) D#/%/'(8!' !@ &/($+ 8!' =S"KK $'3 M/KKK?H3/D/'3/'(
$'3 8'3/D/'3/'( 6/'/#$(8!' !@ /FB/..8%/ ,I ) D#!(/B(8!'
!@ >JA $'3 &8(!B1!'3#8$) $'(8!F83$'( $'3 $'(8H
#
ACS Paragon Plus Environment

Page 11 of 13
ACS Chemical Neuroscience
=N) v l X `;) O`?) PE\\V =N) v l ZP `;) Z`?) PEWV =N) v l VE^ `;)
.!+%/'( A$. (1/' #/&!%/3 $'3 383 $;/!(#!D/ 38.(8++$(8!'
1
2
3
4
5
6
7
8
A8(1 (!+"/'/ $'3 3#8/3 B!&D+/(/+C) A18B1 A$. "./3 $. ."B1
@!# (1/ '/F( .(/D A8(1!"( D"#8@8B$(8!'E 71/' (1/ B#"3/
8'(/#&/38$(/. [$ =WE[ 6) WEPV &&!+? $'3 P$ =WE\ 6) WE\P
&&!+? A$. 38..!+%/3 8' 3#C &/(1$'!+ =[ &+? A18B1 A$.
B!!+/3 (! dS) A$. $33/3 !38"& (#8$B/(!FC 5!#!1C3#83/
= 7A4) ZWW &6) WEPV &&!+?)71/ #/$B(8!' &8F("#/ A$. 0/D(
@!# .(8##8'6 $5!"( O 1 "'3/# '8(#!6/' $(&!.D1/#/E A@(/# (1/
B!&D+/(8!' !@ (1/ #/$B(8!' =&!'8(!#/3 5C 7QS?) .!+%/'(
A$. (1/' #/&!%/3) $'3 (1/ B#"3/ A$. 38+"(/3 A8(1 A$(/# =[
&Q? $'3 #/.83"/ A$. /F(#$B(/3 8'(! b(IAB =Ot \ &Q?E 71/
B!&58'/3 !#6$'8B D1$./ A$. A$.1/3 A8(1 A$(/# =Z t [ &Q?
$'3 5#8'/ =Z t [ &Q?E 71/ !#6$'8B +$C/#. A/#/ B!++/B(/3)
3#8/3 !%/# $'1C3#!". J$O IP $'3 /%$D!#$(/3 8' %$B""& (!
$@@!#3 (1/ B#"3/ D/D(83/ $'3 (1/ #/."+(8'6 #/.83"/ #/H
D#/B8D8($(/3 ".8'6 >SL $'3 `/F$'/) @!++!A/3 5C
(#8("#$(/3 A8(1 38/(1C+ /(1/# (! C8/+3 (1/ B#"3/E 71/ $5!%/
B#"3/ B!&D!"'3 A$. 38..!+%/3 8' >SL =[ &Q? A$. $33/3
7MA =ZWY .!+ 8' >SL?) (1/ #/$B(8!' A$. &!'8(!#/3 5C 7QS
$'3 A$. B!&D+/(/ A8(18' OW &8'E 71/ .!+%/'( A$. (1/'
#/&!%/3 $'3 (1/ #/.83"/ (#8("#$(/3 A8(1 38/(1C+ /(1/# (!
C8/+3 B#"3/ B!&D!"'3. HH&E 71/ $5!%/ B#"3/ B!&D!"'3.
HH& A/#/ D"#8@8/3 ".8'6 $ #/%/#./HD1$./ =,R? ./&8H
D#/D$#$(8%/ `RQS !' SZX B!+"&' $( PW dSE
Z`?) [EV\ =3) v l O `;) [`?) [EOP =33) vl Z[E^) ZPEX `; Z`?) [EZW
=N) v l ZPE^ `;) O`?) OEXX =33) vl ZP) Z[E^ `; Z`? OEVZ =33) v l
ZP) Z[E^ `;) O`?E Z[S JL, =>L IH3^) ZWW L`;? u ZVZEZ)
Z\XEP) Z\^EZ) ZP\EZ) ZPPEZ) Z[PEO) Z[ZE^) Z[WE^) ZOVEW) ZZ]EV) ZZ]EW)
ZZ^EO) ZZ\E[) ZZ\EZ) \XE[) \[EV) \OEW) P\EW) [^E[E `,L =b KHL ?U
+OL B$+B"+$(/3 @!# SO[`OVJPI^ iLe`je P\\EZ]O\) !5./#%/3
P\\EZ]O[E
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
A XN)A4\0 NXG4\G4
(11%8.,*Y )*3%8!".,%*E w71/ ."DD!#(8'6 K'@!#&$(8!'
$#/ $%$8+$5+/ @#// !@ B1$#6/ %8$ (1/ K'(/#'/( $(
1((DUggD"5.E$B.E!#6Ex
AW4=X; )GTX;$A4)XG
N%88&:1%*2,*Y A(.+%8
$
%
%
A(.+%8 N%*.8,-(.,%*:
FVFVJ XV:V %.$ IVWV $(1)'.($ *-( @/+Z(,*V FVFVJ XV:V %.$
eVKV 10.*-(1)f($ *-( ,+&@+3.$1J FVFV 3.$(/*++Y *-(
@-+*+@-01),%9J ;)+@-01),%9J %.$ ). ,(9939+ 1*3$)(1 3.$(/
*-( 13@(/6)1)+. +2 IVWV FVFV %.$ IVWV g/+*( *-(
&%.31,/)@* g)*- ).@3*1 2/+& %99 *-( %3*-+/1V
S!&D!"'3 6U Z`JL, =>L IH3^) PWW L`;?U u ZPE[\ =.) Z`?)
XE]P =.) Z`?) XEVZ =3) vl X `; [ `?) XEZV =( vl ZZEO `; Z`? XEW[
=.) O `?) VE[O =.) Z`?) ^E^PH^E\^ =& Z`?) ^EPZ =N) vl X`;) Z`?
PE^WHPE\\ =&)Z`?) [E^\H[E\\ =&) [`?) [EO[H[EW\ =&)[`?) OE]PH
OEXX =&) Z`?E Z[S JL, =>L IH3^) ZWW L`;? u Z^]EW) Z^^EW)
ZP\EZ) ZP[E^) Z[[EX) ZO]EX) ZO]EW) ZZ]EO) ZZ^E]) ZZ\E]) ZZ\E\) [PE\E
`,L =b KHL ?U +OL B$+B"+$(/3 @!# SZ^`OOJ\IP iLe`je
[PXEZ^^) !5./#%/3 [PXEZ^^E
G%.&:
71/ $"(1!#. 3/B+$#/ '! B!&D/(8'6 @8'$'B8$+ 8'(/#/.(E
0\0)NA4)XG
>/38B$(/3 (! R#!@E ,!'$+3 7E ,$8'/. !' (1/ !BB$.8!' !@ 18.
^W(1 58#(13$CE
Z
S!&D!"'3 GU `JL, =>L IH3^) PWW L`;?U u ZPEO^ =.)
Z`?) XE]^ =.)Z`?) XEVX =3) v l ]E^ `;) Z`)?) XE^^ =3) v l XEP `;)
Z`?) XE\[ =3) v l VEO `;) Z`?) VE]^ =.) Z`?)VE[W =.) Z`?) ^E^O =3)
v l XEW `;) Z`?) ^E\\ =3) v l OEW `;) Z`?) ^EPO =N) v l X `;) Z`?
PEVO =N) v l Z[E^ `;) Z`?) PE[X =N) v l ZP `;) Z`?) [E\X =.) )[`?)
[EW[ =N) v l Z\EO `;) Z`?) OE]PHOEXOZ =&) O`? OEVO =N) v lZP `;)
Z`?E Z[S JL, =>L IH3^) ZWW L`;? u Z^]EW) Z^^EW) Z\XE^)
Z\XE[) Z\XEW) ZP\EZ) ZP[E^) Z[[EX) ZO]EX) ZO]EZ) ZZ]EZ) ZZXEP) ZZXE[])
ZZ\E]) ZZ\E\) \ZE]) [PE\) OVEOE `,L =b KHL ?U +OL B$+B"+$(/3
@!# SZX`OPJ\I^ iLe`je PW^EZVOZ) !5./#%/3 PW^EZVWXE
ANZGX]>\0C$\G4
_/ (1$'0 R#!@E SE JE ,E ,$! M, @!# B!'.($'( ."DD!#( $'3
/'B!"#$6/&/'() vJSA ,) A$#'$v$C$'(8 M/++!A.18D) (1/
>/D$#(&/'( !@ B8/'B/ $'3 7/B1'!+!6C => 7?) 9!%/#'&/'( !@
K'38$ =9#$'(U > 7g vMgS AHWOgOWZ\HOWZ^?) 1/801 $N#
Q$5!#$(!#C
= Q?)
KSL HvJSA ,)
>/D$#(&/'(
!@
48!(/B1'!+!6C) 9!%(E !@ K'38$ =>47g2JH`4HJSH 4gP\Z\?)
S!"'B8+ !@ B8/'(8@8B y K'3".(#8$+ ,/./$#B1 =S K,? @!# .("3/'(
@/++!A.18D @!# @8'$'B8$+ ."DD!#( $'3 R#!@E ,E 2$#$3$#$-$' =KK B)
K'38$? @!# 1/+D 8' K7S &/$."#/&/'(.E
S!&D!"'3 HU Z`JL, =>L IH3^) PWW L`;?U u ZPEZV =.) Z`?)
XE]W =.) Z`?) XEVO =.) Z`?) XE\P =( vl ZZEO `; Z`?) VE[V =.) Z`?)
VEO^HVEOO =&) O`?) ^E]OH^EXP=& Z`?) ^EXO=N) vlVE^ `;) Z`?)
^E^O =3) vlX `;)Z`?) ^E\^ =3) vlO `;) Z`?) ^E[] =N) vlX `; Z`?)
PEWPH[E]P =&) [`?) [EOVH[EZO =&) \`?) OEP^HOE[X =&) O`?E Z[S
JL, =>L IH3^) ZWW L`;? u Z^\EX) Z\XE[) Z\XEW) Z\^EZ) ZP\EZ)
ZP[E^) Z[PE[) Z[ZE^) Z[WE^) ZO]E\) ZZ]EW) ZZ]EW) ZZXEPZ) ZZVE^) ZZ\EX)
ZZ\EP) ZZ\EO) \XEO) PPEV) [PEZ) O\E\c `,L =b KHL ?U +OL
B$+B"+$(/3 @!# SOZ`OPJPIP iLe`je []VEZXV) !5./#%/3
[]VEZXVE
;\T\;\GN\
ZE
R#8'B/) LEc 4#CB/) ,Ec A+5$'/./) bEc _8&!) AEc ,85/8#!)
_Ec M/##8) SE RE 71/ 6+!5$+ D#/%$+/'B/ !@ 3/&/'(8$U $
.C.(/&$(8B #/%8/A $'3 &/($$'$+C.8.E A6L)*!+*#, P*+*&-
GT6H) Q) ^[HV\ /OE
OE _$+.1) >E LEc /+0!/) >E vE >/B8D1/#8'6 (1/ &!+/B"+$#
5$.8. !@ &/&!#C @$8+"#/ 8' A+;1/8&/#o. 38./$./E 2*7#"&
GTT&) DD) ZXZHZ][E
[E ,/8(;) SEc 4#$C'/) SEc L$C/"F) ,E bD83/&8!+!6C !@
A+;1/8&/# 38./$./E 2%-? <*:? 2*7#"6E GT66) R) Z[VHZ\OE
PE A..!B) AE .E OWZX A+;1/8&/#o. 38./$./ M$B(. $'3 M86"#/.E
A6L)*!+*#, P*+*&- GT6U) SD) VWZHVWZE
\E ,$-$./01$#) *Ec S1$0#$5$#(8) LEc 9!%8'3$#$-") 7E
M"'B(8!' $'3 (!F8B8(C !@ $&C+!83 5/($ $'3 #/B/'(
(1/#$D/"(8B 8'(/#%/'(8!'. ($#6/(8'6 $&C+!83 5/($ 8'
Z
S!&D!"'3 &U `JL, =>L IH3^) PWW L`;?U u ZPEW^ =.)
Z`?) ]EWZ =3) vlX `; Z`?) XEXXHXEVP =&) P`?) VE[\^ =.) Z`?)
VEO\ZHVEOZ[ =&) Z`?) VEWX =3) vl^EX `;)Z`?) ^E]W =3) vlX `;)
Z`?) ^EXZ =3) vlVE^ `; Z`?) ^E\X =33) vl \E^) PEW `; O`?) ^EPO
ACS Paragon Plus Environment

ACS Chemical Neuroscience
Page 12 of 13
A+;1/8&/#o. 38./$./E ()*+? ("++7&? GT6O) AS) Z[P[PH
Z[P\WE
^E >!5.!') SE LE) R#!(/8' @!+38'6 $'3 &8.@!+38'6E 2%-7#*
GTTHV DIB) XXPHX]WE
VE ,!..) SE AEc R!8#8/#) LE AE R#!(/8' $66#/6$(8!' $'3
'/"#!3/6/'/#$(8%/ 38./$./E 2%- H*9? GTT&) SC1 ZWHZVE
XE S18(8) MEc >!5.!') SE LE) R#!(/8' L8.@!+38'6) A&C+!83
M!#&$(8!') $'3 `"&$' >8./$./U A "&&$#C !@
R#!6#/.. I%/# (1/ Q$.( >/B$3/E A&&7 <*: !"')*+E
GT6RV TB) OVH^XE
B+/#!.8. 8' (1/ Q$.( >/B$3/E ()*+? <*:?) GT6W) SSQ) ZOOZp
Z[OO
1
2
3
4
5
6
7
8
O[E Q//) Ec T1/'6) tEc *#8.1'$&!!#(1C) vEc $%/+8/@@) LE 9Ec
R$#0) `E LEc 4#/'3/#) vE ,Ec *8&) vE `Ec >/##8B0) vE Ec
*!B18) AEc Q//) `E vEc *8&) SEc ,$&$&!!#(1C) AEc 4!A/#.)
LE 7Ec Q8&) LE `E) ,$(8!'$+ 3/.86' !@ $ .(#"B("#$+
@#$&/A!#0 A8(1 D!(/'(8$+ "./ (! 3/%/+!D B1/&8B$+
#/$6/'(. (1$( ($#6/( $'3 &!3"+$(/ &"+(8D+/ @$B/(. !@
A+;1/8&/#o. 38./$./E 5? A+? ()*+? ;"' GT6&V SUB) O]]H[ZWE
OPE *"&$#) Ec `/''8'6H*'/B1(/+) AEc L$6;!"5) LEc
`$&8+(!') AE >E R/D(83!&8&/(8BH4$./3 L"+(83!&$8'
7$#6/(8'6 I@@/#. S#8(8B$+ b%$+"$(8!' !@ A5/($ (#"B("#/
$'3 7!F8B M"'B(8!'E 5? A+? ()*+? ;"' GT6U) SDC) ^\^OH
^\VPE
O\E 4/B0) LE _Ec >/##8B0) vE Ec */##) ,E AEc I1) E 4Ec S1!) _E
vEc Q//) E vEc v8) {Ec `$') vEc 7/1#$'8) TE AEc "1) JEc *8&)
Ec Q$#./') E >Ec *8&) *E Ec Q//) vE {Ec ,"!(!+!) 4E 7Ec Q8&)
LE `E (#"B("#/H&/B1$'8.&H5$./3 /'68'//#8'6 !@
B1/&8B$+ #/6"+$(!#. ($#6/(8'6 38.(8'B( D$(1!+!68B$+
@$B(!#. 8' A+;1/8&/#o. 38./$./E 2%-? ("++7&? GT6P) R)
Z[ZZ\E
O^E L"(1"#$-) 4Ec Q$C/0) Ec 4$+$-8) E JEc 7#8%/38) 2Ec KC/#) RE
*E L"+(8D+/ @"'B(8!' @+"!#/.B/8' D#!5/ D/#@!#&. &/($+
B1/+$(8!') 38.$66#/6$(8!') $'3 &!3"+$(8!' !@
$66#/6$(/3 A5/($ $'3 A5/($HS" B!&D+/FE A(; ()*+?
2*7#",'!? GT6O) B) ZXXWHZX]ZE
OVE 4$##!A) SE vEc T$6!#.08) LE 9E !+"(8!' .(#"B("#/. !@ 5/($
D/D(83/ $'3 8(. B!'.(8("/'( @#$6&/'(.U #/+$(8!' (!
$&C+!83 3/D!.8(8!'E ;'!*&'* 6WW6) IAU) ZV]HZXOE
OXE ,$-$./01$#) *Ec "#/.1) E JEc L$'-8(1$C$) ,Ec
9!%8'3$#$-") 7E ,$(8!'$++C 3/.86'/3 D/D(83!&8&/(8B
&!3"+$(!#. !@ $5/($ (!F8B8(C 8' A+;1/8&/#o. 38./$./E ;'!?
<*N? GT6O) A) XZ[]E
O]E ,$-$./01$#) *Ec J$#$C$'$.A$&C) JEc L8.1#$) REc "#/.1)
E JEc L$'-8(1$C$) ,Ec 9!%8'3$#$-") 7E C'(1/.8. !@
`C5#83 SCB+8B R/D(!83. $'3 K3/'(8@8B$(8!' !@ A"(!D1$6C
b'1$'B/#E ()*+N67,')*+E GT6&) RQ) O\H[WE
[WE ,$-$./01$#) *Ec L$31") SEc 9!%8'3$#$-") 7E J$("#$+
7#8D/D(83/H4$./3 K'1858(!# !@ L"+(8@$B/(/3 A&C+!83
5/($ 7!F8B8(CE A(; ()*+? 2*7#",'!? GT6P) R) Z[WWHZ[ZWE
[ZE R8B0$#() QEc 2$.N"/;H !+(/#!) vE LEc L$#6!+8'$) AE 9`*
R/D(83/ $. $ J$("#$+ L!3"+$(!# !@ L"+(8D+/ S/++"+$#
R$(1A$C. 8' 08' ,/6/'/#$(8!'E !"+*9 <*, F&- GT6O)
ICSA) ^PXZWXE
[OE ,/C58/#) *Ec AC$+$) Ec A+8/.) 4Ec ,!3#86"/.) vE 2Ec 4".(!.
,!3#86"/;) Ec Q$ R/''$) 9Ec S!++8') MEc 9!&/.) SE LEc
`"#/$") SEc M$++/#) RE M#// "D/#!F83/ 8. $' K'(/#&/38$(/
8' (1/ R#!3"B(8!' !@ `OIO 5C S!DD/#=K?HA5/($ R/D(83/
$'3 IOE A&$*3? ()*+? F&-? V9? GT6P) AA) ZWX\HZWX]E
[[E S1/86'!') SEc 7!&$.) LEc 4!''/@!'(H,!"../+!() >Ec
M$++/#) REc `"#/$") SEc S!++8') ME IF83$(8%/ .(#/.. $'3 (1/
$&C+!83 5/($ D/D(83/ 8' A+;1/8&/#o. 38./$./E <*9"G !"6
GT6U) SD) P\WHP^PE
9
]E `$&+/C) KE _E 71/ $&C+!83 5/($ D/D(83/U $ B1/&8.(o.
D/#.D/B(8%/E ,!+/ 8' A+;1/8&/#o. $'3 @85#8++8;$(8!'E
()*+? <*:? GT6G) SSI) \ZPVH\Z]OE
ZWE `$($8) vEc L!(8/8) QEc L$#6"+8/.) >E A'$+C;8'6 A&C+!83
4/($ A66#/6$(/. A8(1 $ S!&58'$(!#8$+ M+"!#/.B/'(
L!+/B"+$# /'.!#E 5? A+? ()*+? ;"' GT6R) SUQ) OZ[^HOZ[]E
ZZE !A$3/) ,E MEc v$1') 7E ,E //3H8'3"B/3 $BB/+/#$(8!' !@
$&C+!83H5/($ &/38$(/3 '/"#!(!F8B8(C 8' %8%!E 2%-?
("++7&? GT6R) T) \ZOE
ZOE `"C) RE >Ec 2"!'6) rE 2Ec Q$ R/''$) 9Ec M$++/#) REc Q8) LE
E K&D$B( !@ S"=KK? 48'38'6 !' (#"B("#/. $'3 >C'$&8B.
!@ A5/($PO L!'!&/# $'3 >8&/#U L!+/B"+$# >C'$&8B.
("3CE A(; ()*+? 2*7#",'!? GT6P) R) Z[PXHZ[^[E
Z[E `$#3C) vEc /+0!/) >E vE 71/ $&C+!83 1CD!(1/.8. !@
A+;1/8&/#o. 38./$./U D#!6#/.. $'3 D#!5+/&. !' (1/ #!$3
(! (1/#$D/"(8B.E ;'!*&'* GTTG) IQR) [\[H[\^E
ZPE 9$66/++8) bEc *!;+!A.08) `Ec 2$+/'.8') >Ec 2$+/'.8') 9E
S!DD/# 1!&/!.($.8. $'3 '/"#!3/6/'/#$(8%/ 38.!#3/#.
=A+;1/8&/#o.) D#8!') $'3 R$#08'.!'o. 38./$./. $'3
$&C!(#!D18B +$(/#$+ .B+/#!.8.?E ()*+? <*:? GTTP) SCB)
Z]]\HOWPPE
Z\E S"$3#$3!) AEc ,!-!) AE KEc _/++.) 9Ec `$C/.) vE >Ec S!".8')
E REc ,"&./C) _E QEc A(("B0.) IE SEc M#$'0+8') Ec
Q/%!'/') AE QEc */'.+/#) 7E _Ec >8'0!%$H*!.(!%$) AE 7E)
71/#$D/"(8B ($#6/(8'6 !@ (1/ J,MO $'3 *bARZ
D$#('/#.18D 8' B1#!'8B 38./$./.E 2%- <*: P#7$ P!,'":?
3!8U ZWEZW[Xg.PZ\V[HWZXHWWWXHFE
Z^E _C..HS!#$C) 7E K'@+$&&$(8!' 8' A+;1/8&/# 38./$./U
3#8%8'6 @!#B/) 5C.($'3/# !# 5/'/@8B8$+ #/.D!'./z 2%-
H*9? GTTP) SI) ZWW\HZWZ\E
ZVE `/'/0$) LE 7Ec LBL$'".) ,E LEc Q$(;) bE K'@+$&&$.!&/
.86'$++8'6 8' 5#$8' @"'B(8!' $'3 '/"#!3/6/'/#$(8%/
38./$./E 2%- <*: 2*7#",'! GT6UV SQ) ^ZWH^OZE
ZXE L$.(#!/'8) >Ec *13!"#) IE LEc >/+%$"F) bEc J!+;) vEc
I+./') 9Ec 4/#B1(!+3) JEc S!(&$') SEc `/B1() E LEc
S!+/&$') RE >E J"B+/$# 5"( '!( &8(!B1!'3#8$+H/'B!3/3
!F83$(8%/ D1!.D1!#C+$(8!' 6/'/. $#/ $+(/#/3 8' $68'6)
&8+3 B!6'8(8%/ 8&D$8#&/'() $'3 A+;1/8&/#o. 38./$./E
A6L)*!+*#, P*+*&-E GT6RV SU) \ZWH\Z]E
Z]E ,$-$./01$#) *Ec L/1($) *Ec 9!%8'3$#$-") 7E `C5#83
L"+(8@"'B(8!'$+ L!3"+$(!#. K'1858( L"+(8@$B/(/3 A5/($
7!F8B8(C $'3 R#/%/'( L8(!B1!'3#8$+ >$&$6/E A(;
()*+? 2*7#",'!? GT6U) Q) ZP[OHZPPWE
OWE ,$-$./01$#) *Ec 9!%8'3$#$-") 7E S"##/'( D#!6#/..)
B1$++/'6/. $'3 @"("#/ D#!.D/B(. !@ 38$6'!.(8B $'3
(1/#$D/"(8B 8'(/#%/'(8!'. 8' A+;1/8&/#o. 38./$./E <;(
A9:? GT6U) T) O[VXWHO[XWPE
OZE *$##$') bEc L/#B0/') LEc >/ (#!!D/#) 4E 71/ $&C+!83
B$.B$3/ 1CD!(1/.8. @!# A+;1/8&/#o. 38./$./U $' $DD#$8.$+
@!# (1/ 3/%/+!D&/'( !@ (1/#$D/"(8B.E 2%-? <*:? P#7$
P!,'":? GT66) SC) ^]XHVZOE
OOE $%/+8/@@) LE 9Ec J$&) 9Ec *$'6) vEc Q//) `E vEc Q//) LEc
Q8&) LE `E >/%/+!D&/'( !@ L"+(8@"'B(8!'$+ L!+/B"+/.
$. R!(/'(8$+ 71/#$D/"(8B S$'383$(/. @!# A+;1/8&/#o.
>8./$./) R$#08'.!'o. >8./$./) $'3 A&C!(#!D18B Q$(/#$+
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
[PE 4$#'1$&) *E vEc 4".1) AE KE 48!+!68B$+ &/($+. $'3 &/($+H
($#6/(8'6 B!&D!"'3. 8' &$-!# '/"#!3/6/'/#$(8%/
38./$./.E ()*+? ;"'? <*:? GT6&) DU) ^VOVH^VP]E
[\E */DD) *E RE 48!8'!#6$'8B B1/&8.(#C !@ A+;1/8&/#o.
38./$./E ()*+ <*: GT6G) SSI) \Z][H\O[]E
[^E S$5$++/#!) AE 4Ec 7/#!+HI#3$;) QEc b.D$#6$#!) AEc
2$;N"/;) 9Ec J8B!+$.) bEc $5$(/) ,Ec 9$&/;) RE
`8.(838'/H,8B1 I+86!D/D(83/. 7! Q/../' S!DD/#H
L/38$(/3 A&C+!83H5/($ 7!F8B8(CE ()*+? V7#? 5? GT6P) II)
VO^XHVOXWE
[VE _18(/1!#'/) 7E vEc B1$D/#) ME N"$#/HD+$'$# S"=KK?
380/(8&8'$(/ B!&D+/F/. 8' +$B(83/ D!+C&/#8;$(8!'E
F&"#$E ()*+E GT6H) AI) Z[^ZOHZ[^OOE
!
ACS Paragon Plus Environment

Page 13 of 13
ACS Chemical Neuroscience
[XE 9#$5!) vE bEc S1#8.&$') LE AEc _/55) QE LEc 4$+3A8') LE
PPE R$++!) E REc >8L$8!) vEc S!!0) AEc J8+..!') 4Ec v!1'.!') 9E
2E _E L/B1$'8.&. !@ ($" $'3 A5/($H8'3"B/3
/FB8(!(!F8B8(CE 4#$8' ,/.E GT6PV Z^[P) ZZ]HZZ[ZE
P\E R+$.B/'B8$H28++$) 9Ec R!'B/) AEc S!++8'6A!!3) vE MEc
A#/++$'!Hv8&/'/;) LE vEc T1") tEc ,!6/#.) vE 7Ec
4/($'B!"#() KEc v!./H{$B$&$') LEc R/##C) 9E) `861H
#/.!+"(8!' $'$+C(8B$+ 8&$68'6 $'3 /+/B(#!' 1!+!6#$D1C
!@ &$6'/(8(/ D$#(8B+/. 8' $&C+!83 B!#/. !@ A+;1/8&/#o.
38./$./E ;'!? <*N? GT6P) B) OPXV[E
P^E S!++8'6A!!3) vE MEc S1$'3#$) Ec >$%83.!') LEc
L801$C+!%$) AEc b.08') 7Ec >!5.!') vEc M!#3/#) vEc 4$(8B1)
SE) ROHWWZU `861H#/.!+"(8!' &$6'/(8B #/.!'$'B/
8&$68'6 (! N"$'(8@C #/+$F$(8!' D$#$&/(/#. 8'
A+;1/8&/#o. 5#$8' (8.."/E A6L)*!+*#, P*+*&-E GTTU) D)
[\^H[^^E
vE R1!(!B1/&8B$+ #/$B(8%8(C !@ (1/ 8#!'=KKK? B!&D+/F !@ $
&8F/3H3!'!#) $+D1$H1C3#!FC $B83HB!'($8'8'6 B1/+$(/
$'3 8(. 58!+!68B$+ #/+/%$'B/ (! D1!(!$B(8%/ &$#8'/
.83/#!D1!#/.E F&"#$E ()*+E GT6&) AU) \VXZH\VXVE
1
2
3
4
5
6
7
8
[]E L/'$) Ec L8#$(.) AEc S$5$++/#!) AE 4Ec 9"8#$3!) 9Ec
4$##8!.) QE AEc 7/$() E vEc ,!3#86"/;H $'(8$6!) QEc !3"D/)
LEc 9$&/;) RE >#$.(8B b@@/B( !@ (1/ R/D(83/ /N"/'B/ !'
(1/ S!DD/#H48'38'6 R#!D/#(8/. !@ 7#8D/D(83/. $'3 (1/
b+/B(#!B1/&8B$+ 4/1$%8!"# !@ 71/8# S!DD/#=KK?
S!&D+/F/.E ()*+? V7#? 5? GT6U) ID) \Z\[H\Z^OE
PWE ,!5/#() AEc Q8") {Ec J6"C/') LEc L/"'8/#) 4E ,/6"+$(8!'
!@ B!DD/# $'3 8#!' 1!&/!.($.8. 5C &/($+ B1/+$(!#.U $
D!..85+/ B1/&!(1/#$DC @!# A+;1/8&/#o. 38./$./E A''
()*+ <*,? GT6O) DT) Z[[OHZ[[]E
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
PZE Q8") TE rE S1/&8B$+ 8'.861(. 8'(! 68'./'6 $. $ #/.!"#B/
@!# '$("#$+ $'(8!F83$'(.E ()*+? <*:? GT6G) SSI) [[O]H[[\\E
POE t8/) vEc B1$8B1) *E LE ,/H/%$+"$(8!' !@ (1/ O)OH38D1/'C+H
ZHD8B#C+1C3#$;C+ @#// #$38B$+ =>RR`? $..$C @!#
$'(8!F83$'( $B(8%8(CE 5? A$#!'? W""9 ()*+? GT6&) BI) PO\ZH
PO^WE
P[E 48.A$.) AEc *"#0"(/) REc $+//&) Ec v$'$) 4Ec L!1$D$(#$)
Ec L!'3$+) REc A3$0) AEc 91!.1) Ec $1$) AEc 41"'8$) >Ec
48.A$.) E SEc 91!.1) E J!%/+ 1/F$D/D(83/ 8'(/#$B(.
A8(1 ("5"+8' $'3 &8B#!("5"+/.) 8'1858(. A5/($
@85#8++$(8!') $'3 .1!A. .86'8@8B$'( '/"#!D#!(/B(8!'E A(;
()*+? 2*7#",'!? GT6O) B) Z[W]HZ[Z^E
PVE 4!"#$..$) LE _Ec L8++/#) QE LE) L/($+ 8&$68'6 8'
'/"#!3/6/'/#$(8%/ 38./$./.E H*-%66"+!',E GT6G) D) VOZH
[XE
PXE 4$#'1$&) *E vEc 4".1) AE KE) 48!+!68B$+ &/($+. $'3 &/($+H
($#6/(8'6 B!&D!"'3. 8' &$-!# '/"#!3/6/'/#$(8%/
38./$./.E ()*+ ;"' <*:? GT6&) DU) ^VOVHP]E
P]E `"'(/#) ,E AEc (!#&) _E QEc S!'/.08) RE JEc B1!/'@8.B1)
LE `E K'$BB"#$B8/. !@ '8(#8B !F83/ &/$."#/&/'(
&/(1!3. 8' 58!+!68B$+ &/38$E A&%6? ()*+? GT6H) TA) Z]\VH
Z]^[E
"
ACS Paragon Plus Environment