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`_; TRANSPERFECT
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`County of Kings, City of New York, State ofNew York
`
`Translation Certificate
`
`1, Philip Schnell, am fluent in the Japanese and English languages and have
`personal knowledge of the facts stated herein.
`Attached hereto is a document entitled “(Jpn. J. Neuropsychopharmacol.) 19 :
`363 (1999)” and a true and accurate translation of “(Jpn J.
`'
`acol.) 19 : 363 (1999)”,from Japanese into English.
`
`
`
`Sworn to before me this
`
`lst day of November 2019
`
`Signature, Notary Public
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`mm
`manners-um
`norm
`Whitman!!!
`mmwauDZ-L
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`Stamp, Notary Public
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`Page 1 0f 3
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`SLAYBACK EXHIBIT 1028
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`Page 1 of 3
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`SLAYBACK EXHIBIT 1028
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`

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`EP$:~J!M~IHI~·Ii (Jpn. J. Neuropsychopharmacol.) 19: 363 (1999)
`
`: .-jl:: BHatt;,ri LlbHAAV
`->CIF •f'r: n:-f'U"-'f1' OG'
`22MAY201J
`
`363
`
`.WfMI.tJ[fitf!lljp,f~ SM-13496 OJ1il1tl~I¥J%
`tl :lli$ fl'l ~HI 51t~€Hfi: 1:: M 9 ofF JlUffllli
`
`Binding affinity of SM-13496 to various receptors
`Receptors
`Ki values (nM)
`
`Ottll~ tr-1-. EHJJl ftl~. lf!Al -JIJ .
`:k!f ffi'J.
`ttii:!t!!i <~> · .ttJ!!i~-liif~m
`1"554-0022 .:k~ilitlt:tt~li B lli Jtl 3-1-98
`
`SM-13496 li~ :htJ.tit }'-!-/~ ~ /tfJH!>J::'CttittD 1-
`=/~ffl~~G~~S . • ~~~-~tfffl!>J::'CtJtiM~~
`tU•J ~ ffl tM~ 1in:: ~ 1r) 4-'i tl ~ ~ "9 ¢.if J:f/. tltftH!tliU~ I: a;
`¢. ~@J. SM-13496 Q)~ft~(J(JtJ.~J)J!~tt~~ 6b\l:
`"9 ¢t:::.~. S~11fl~~if~Jil~!fJ~cQ)-MJ:!i~JIH:"Jir)"t
`Wt itt ~ fi' ·:>t::;. •
`
`l.~~~fiifi~~
`~m~~t.t>1:~-r ¢f.'it.tJtftlttQ)f¥ilili*'tH:~? l
`Fht:::.. "tt.t.b!>. i!illil::t:::.l;t~~~§l.IJlMUU~db~ 1J
`n;.;r:.~~~t.~c~~m~T~:~;.;~~~-~3/G
`t::;.. -fQ)lfi, ~E£-?i§I:J::I'J#7.AU-?.tiLtl:-?iAG.
`~m~a@m-~~:R~G~.-?ffi~~~~"9¢MNMtt
`Hl4l:E"t ¢;: c I:J:: I'J. 4?;~a<J16tl~ 50%1!Ilt!J"t ¢ ~~t•ft
`B'4tQ)iftlll: (IC110) ~;1<6'), Ki= IC501' (1+S/Kd) I:J:: I'J Ki
`t-: iftll:) •
`{ifi H'Hl:l t.., 1::. (S: IJ # /
`2. 77=Jv~ :/ '7 7--EJt;&-&.~t-•l:t>ft ¢~Jil
`1:: 1- DtL1t~i*HVJl~itt.:: CHO IBI'c2 (CHOihDsLMll
`~) ~ KRH 81itillll:@i!ll L,.,, 10 JJ]vl forskolin. iU1HtB'
`4joj, 300nMdopamine~1JD;t'l37"C. 20~~/.:f~-"'­
`:/3/G~. 0.5mMCHsCOONa(pHG.2) ~1JD;t. 1071'
`rlll~~G"t&Jt~.tl::.li6. ~,c.,I:J::I'J~M~'ltllkG~. 8:~
`L,., t::;. cAMP li EIA cAMP assay kit (Amersham)~ Jfl ~l "(
`~);£ L t:::.. Forskolin ~~ cAMP 1::~9 ¢ dopamine 0)~
`~~50%1fiJ!f;!J9¢?1Utdtft~Q)·/l (EC50) HJWGt:::..
`3.MI!~I*!JJ.Itl:!llt o [3H]ACh .irflilfH:M"t ¢~ffl
`7 'l ~-~~~lJ.l.lt (0.3X0.3 uAi) ~ 95%0:ti5%COt
`.iili ~ 'f "t' Tyrode UUIJ ~ 1::!! l!ti L,., • 1 nM PH] choline
`chloride ~l'JS1JJ {..., "'(. 37'C I: 30 71"1 /.:f ~ -"'- :/3 / L,.,
`tO J,lM hemicholinium-3 ~~tl' Tyrode *IIi
`t:::. • .of Q)lfl,
`~"f 3 @I Wash Gt:::.. ;:tu::. 20 mM KCI. ~~t•fttl~.
`1 J,lM quinpirole 'f: JJll;t "'( 37'C I: 10 5.Hifl1' /.:f ~ -"'- :/
`3/G~.-t-O)~.*~mm~~w;t,~ff-?~~.ti'Jn
`7.Afi*ft-?ki{.ti:-?~G. &R\~~.tl::.L,.,t::;.. [3H]AChilt_.
`$= -?Ill rr 0)[3H]tiUtMttt(-? ?!l Jt1 Q) PHJMNMtt+ -?*U
`~Q)[3H]i~UtiZitt) c Gt:::.. jgj K•!ti•Jlr£[3H]ACh iliMI:~"t
`¢ quinpirole Q)ttlJflii!~ 50%11llf!liJT¢11U~ftft~O)·J.i:
`(EC110) ~ JJI:tl L,., t.:.
`
`16~:8J::U::iltll
`SM-13496 O) .~.HII1t~i*1:~"9¢Uift~Untt~~I:;E c
`~t:::.. SM-13496 l:t t-:-1~~ / Dz, tot-=/ 5-HT2A,
`
`Dopamine (DA)
`o.
`Dt
`DA uptake sites
`Serotonin (5-HT)
`5-HTIA
`5 - HT:~A
`5-HT!c
`5-HTs
`5-HT•
`5-HT7
`5-HT uptake sites
`Noradrenaline
`Ul
`<12
`CUA
`we
`~
`~·
`~2
`Muscarine
`M1
`M:~
`Histamine 1 (H,)
`Benzodiazepine
`Opiate
`Sigma
`•> ICc,o vnlue
`
`262
`1.68
`>1000•)
`
`6.75
`2.03
`415
`>1000 •J
`>1000 •J
`0.495
`>1000 •>
`
`47.9
`66.7
`40.7
`10.8
`>1000 •l
`>1000 o)
`>1000 •>
`
`>1000 •>
`>1000 •>
`>1000 •>
`>1000 •>
`> 1000 a)
`>1000 o)
`
`5-HT!A• 5-HT, :BJ::L( 7 }'-! v'Tl) / <l!c ~!fJi*l:~lr\JS
`fifJI.foi1~:if- L,.,~. -'jj. a,, <l2A· D1 :BJ::U: 5-HTrc ~
`~~~~To~B-fJI.~tt~~~~~<.~Q)•Q)•<O)~
`~~~~16t.Mi1~:if-~~~?t.:. il::~.MQ)~#11fl~~
`
`c Q) JU.lh'J\ 6 . 1) SDA ~ t1i M. m. ( risperidone,
`
`olanzapine fJ.C> I:M~ l,.,"'( 5-HTtA~~~~:~"9¢.atl
`IUnttiJf~ If);: c, 2) 5-HTIA• 5-HT7 , Ute 1t~i-$1:~
`9 otlit.#tfni1n!ft6~t\;: c. 3) a • .t:>J:: a 1:: .A :7:::: ;,; H. 1t
`~I* 1:~"9 o $JUili1tJ1MAlJ 1:~"'#1lt 1: at If);: c :Q!:if- ~
`tl~. -'jj, CHOihD:~L a&lfc21::8lr\"(, SM-13496 li
`dopamine I:J::¢ cAMP .MI£ttllfti!l&Jt.\~t'il1l G, ~Q)tf.ffl
`li haloperidol c flll~. risperidone. chlorpromazine !>J::
`U:clozapine l:::~"'il§I'J\?t.:. ;:nl:~t..,. ~'~•i*W.Jtl:
`t.Hto quinpirole I:J::¢ ['H]ACh ia-1!llftitl&.lt\l:t~9
`¢ SM-13496 O)fa~tf}lll;tjjj) <,haloperidol Q) 1/30 W.T
`I: <I;?~.
`J;J.~O)JU~J:: I'J, SM-13496 l:t D2 !>J:.'Ct 5-HTt1t~
`~~~~.ffl~ftG"tm•'lt~m~~~&•-r.o;:e:.a::~.
`a •. H. t>J::U 5-HTtc ~tli*l:::~"toiiitli!~tttJ!iltv)
`;: c:Q\6, ~Q)l'J!aiB~. 'PM111JIWtti11Jff.ffl:8J::'Cti*lll.ft
`Wff=JiliJf~lr\;:cb!:if-'Jf~tl¢. ~SI:::, i$~{.$ Dz~fJ
`t.t>~ftGt::.7t7Jv:J 'J ;.;.-fi~l:::~9¢ff.ffltJfiJ;it,.);:
`~~6.~Q)•~~~-~tfm~a~-ra;¢;:c~~'Jf~
`tl¢.
`
`Page 2 of 3
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`SLAYBACK EXHIBIT 1028
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`

`

`Japanese Journal of Neuropsychopharmacology (Jpn. J. Neuropsychopharmacol.) 19: 363 (1999)
`
`
`
`P-3) Pharmacological characteristics of the novel
`antipsychotic SM-13496: Evaluation of action on various
`receptors in the brain
`
`
`
`
`363
`
` Tomoko HORISAWA, Rie TAGASHIRA, Kazuki
`YABUUCHI, and Yukihiro ŌNO
`First Drug Discovery Laboratory, Sumitomo
`Pharmaceuticals Co., Ltd., 3-1-98 Kasugade-Naka,
`Konohana-ku, Ōsaka-shi 554-0022
`
`
` SM-13496 is a novel antipsychotic that, while having pow-
`erful anti-dopamine action and anti-serotonin action, is charac-
`terized in having extremely weak extrapyramidal side effects
`and central depression side effects. In this paper, we studied
`interactions with brain neurotransmitter receptors in order to
`clarify the biochemical pharmacological properties of SM-
`13496.
`
`
`Method
`1. Receptor bonding experiment
` Bonding affinity to various receptors was evaluated in ac-
`cordance with the conventional method. Specifically, prepara-
`tions of tissue or receptor-expressing cell membranes were
`incubated in a constant-temperature bath together with the test
`compound. Then, the mixture was filtered onto glass fiber filter
`paper by reduced-pressure filtration and the filter paper was
`washed several times with a buffer solution. By measuring
`radioactivity remaining on the filter paper, the concentration
`(IC50) of the test compound at which specific bonding was
`50% inhibited was determined, and the Ki value was calculated
`using the formula Ki = IC50 ∕ (1 + S ∕ Kd) (where S is the ligand
`concentration).
`2. Action in adenylate cyclase coupled reaction system
` CHO cells (CHO/hD2L cells) made to express human D2L
`receptors were suspended in a KRH buffer solution, 10 µM of
`forskolin, the test compound, and 300 nM of dopamine were
`added, and the mixture was incubated for 20 minutes at 37°C.
`0.5 mM of CH3COONa (pH 6.2) was added, the mixture was
`boiled for 10 minutes to stop the reaction, and the supernatant
`was portioned out by centrifugation. The cAMP produced was
`measured using an EIA cAMP assay kit (Amersham). The
`concentration (EC50) of the test compound at which dopamine
`suppression of forskolin-induced cAMP was 50% suppressed
`was calculated.
`3. Action on [3H]ACh release in striatal slices
` Rat striatal slices (0.3 × 0.3 mm thickness) were suspended
`in a Tyrode’s buffer solution with 95% O2/5% CO2 ventilation,
`1 nM of [3H]choline chloride was added, and the mixture was
`incubated for 30 minutes at 37°C. Then, the mixture was
`washed three times with a Tyrode’s buffer solution including
`10 µM of hemicholinium-3. To this, 20 mM KCl, the test com-
`pound, and 1 µM of quinpirole were added, and the mixture
`was incubated for 10 minutes at 37°C. Then, an ice-cooled
`buffer solution was added, the mixture was filtered by reduced-
`pressure filtration onto glass fiber paper filter paper, and the
`reaction was stopped. The [3H]ACh release rate was consid-
`ered to be the [3H] radioactivity in the filtrate ∕ (the [3H] radio-
`activity in the filtrate + the [3H] radioactivity on the filter pa-
`per). The concentration (EC50) of the test compound at which
`quinpirole suppression of high-K+-stimulated [3H]ACh release
`was 50% suppressed was calculated.
`
`
`
`Results and discussion
`The bonding affinity of SM-13496 to various receptors is
`summarized in the table. SM-13496 showed high bonding
`affinity to dopamine D2, serotonin 5-HT2A, 5-HT1A, 5-HT7, and
`adrenaline α2C receptors. On the other hand, bonding affinity to
`α1, α2A, D1, and 5-HT2C receptors was relatively weak, and no
`bonding affinity was shown to many other receptors. Moreover,
`a comparison with other antipsychotics showed that 1) bonding
`affinity to 5-HT2A receptors was high, similarly to that of
`SDA-based antipsychotics (risperidone, olanzapine, etc.), 2)
`bonding affinity to 5-HT1A, 5-HT7, and α2C receptors was high,
`and 3) affinity to α1 and histamine H1 receptors was extremely
`low by comparison with the other drugs. On the other hand, in
`CHO/hD2L cells, SM-13496 was antagonistic to cAMP produc-
`tion suppression reactions caused by dopamine, and this action
`was equivalent to that of haloperidol and stronger than that of
`risperidone, chlorpromazine, and clozapine. In contrast, the
`antagonistic action of SM-13496 on [3H]ACh release suppres-
`sion reactions caused by quinpirole in striatal slices was weak,
`one-thirtieth or less that of haloperidol.
` These results suggest that SM-13496 improves schizophren-
`ic symptoms via D2 and 5-HT2 receptor blocking action and
`that its circulatory system and central depression side effects
`and body weight increasing action are weak because its bond-
`ing affinity to α1, H1, and 5-HT2C receptors is low. Furthermore,
`the results suggest that the drug’s extrapyramidal side effects
`are gentle because its action on the acetylcholine nervous sys-
`tem via striatal D2 receptors is weak.
`
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