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`~ ca ndinav i an U niversity Press, Oslo - Stockholm
`
`VOL 72.
`
`Dr. Reddy's Laboratories, Ltd., et al.
`v.
`Galderma Laboratories, Inc.
`IPR2015-__
`Exhibit 1032
`
`

`
`.. •
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`I
`
`..
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`...... ,~ ...... .,,,'-..,.. .... •••'-'•• ...
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`I
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`\cand inavian U nivers ity Press, Os lo- Stockholm
`
`VOL. 72. 1992. No.3
`
`Exh. 1032
`
`

`
`Acta D enn Vc nc reo l ( Stock h ) 1992: 72 : J7S- J7lJ
`. s·
`Effect of Doxycycline on the Generation of Reactive Oxygen Specle ·
`A Possible Mechanism of Action of Acne Therapy with Doxycycline
`
`1-IIROI-I I KO 1\ K A M /\T SU . M A KI A SA D /\ . . JINR.O KOM U R A. Y AS UO A SA D A . and YUKIE NIWA
`fletwrllll<'lll of IJ emullo!og\' , Kunwi f\olcdicul U11in·rsitv, 0.111k11 570. und 1 N ilV<I fn stitllll' j i>r fn!lllltllolo g \'. 1\ocl!i 787--03 . .Iot"'"
`
`1
`
`On the hasis of a r·el·ent r·eport tlrat minol·ycline is d'fective in
`IIre treatmcnt of acnc inflammation hy acting dir·cctly as an
`antioxidant on infiltrating ncutrophils, we investigated whcther
`doxycyclinc might also be capahlc of r·edudng thc genenrtion uf
`reactive oxygen species, using human neutmphils and a ccll-
`frce, xanthinc-xanthine oxidase system. The spccics investi-
`gated ar·e Superoxide nrd ical anion (0 ~ ), hydr·ogcn per·uxidc
`m,O,) and hydroxyl radkal (OH · ). Doxycycline signilicantly
`r·educed the Ieveis of 0 ~ . H,O, and OH· generated hy hoth
`systems. Our· results scem tu suggest that thc clinical cll'cctivc-
`ness of doxycycline in thc treatmcnt of acne intlammation is due
`partly tu its antioxidant cffect on ncutmphils. Kcy words: Anti-
`oxiduni action; N eutropllil; Tetracyclines.
`( /\cce pt ed Se pt ember 2.>. JlJl)l_)
`
`/\ cta D en n V enereol ( Stockh) 1992: 72 : 17S- 179.
`
`II. 1\ kamat su . D epartment of Dcrmatology. Kan sai M edical
`U ni ve rsit y. I Fumi zono-cho . Morig uc hi -s hi . ü saka 'i70 . .Ia-
`pan
`
`Tct r;JcyL·Iin es. such as minocyc lin e and do xycyclin e. are an
`ciTecti ve tre; Jtm ent fur ac ne v ul gari s. W c ha vc re ce ntl y re-
`port ed that minocyc li ne inhihits neutrophil reacti ve oxyge n
`species (ROS) gL·ncration . w hich support s th c hypoth es is that
`th e age nt is d fecti vc in acn e trc atm c nt not o nl y h y redu cin g
`lh L· llU lllhc rs o f I'J 'O{Jinnihoctl'ritllll lli'JH'.I' (/'. tti'Jil'.l) . hut ;dso h y
`inhihiting nc ulrophi l-ge ncr;Jl ed RO S w hi ch indu ce a chc m ical
`in su ll to lhc inll'g rit y of folli cuLir epilh elium in <Jene ( I ). W e
`h;1vc morc I;Jl ely found th at rc ducc d Ie ve ls of linolcic acid i n
`<tc ne con! c donc;, coillJ1<1rahlc with normal hair fo ll iclc s, w hi ch
`d k cti ve ly inhihits nc ulruphil ROS gc nc rat ion . contrihute s to
`;1n ex;,ccrh;Jtion of ac ne infl;umnatio n ( 2).
`/\ lthoug h do xycyclin e ha s Ilee n report ed to ha ve a ROS -
`redu cin l'. d lcc l (.'l ) . lhc cflcct on th e lcVL' Is of all kinds of ROS
`ge n e ral ~ d hoth h y neulrophil s and in lhc x anlhine-x anthin c
`o xid;1se sys tem ha s nol yc l Ilee n examin c d .
`l n lh e present stud y. we invesli gat ed w helhc r do xycyclin e
`mi ghl he e< ipahlc of rcdu cin g thc gc neration of ROS . includin g
`supcro xid e r;1dical anion (0, ), h ydrogt'- n pero xiLic (II , OJ and
`h ydrox yl radi c; d (Oll·). usin g human ncutrophils ;u!d a ce ll -
`frec. xanthin c-x anlhin e o x idase sys te n1. Th e res ult s reve alcd
`thal do xycyc lin c does in L1 cl dlccli ve ly inhihil the Ie ve ls of all
`k ind s of RO S genc r;il cd in ho lh sys tem s
`
`M/\ 11-:RI /\ I .S /\ N D M I TIIOD S
`( 'ill'llll m ls: l )o xycvl'lin c 1 L1it n l'li;.-n l'h;mnaCL' llliL·;Ih l.td . Japan) w;1'
`:tdd n l tu th e lull nw in ~ ncutrophil fun c tio11 <ts:-.<t y -.. ys iL' Ill \ in COilL"L' Illrtt -
`1 '" " ' <> I ll .llll'i. ll .ll'i . ll .'i. 'i '" " 1 .'ill 1' ~./ 1111 .
`N nilrt}/)ltd l"'l'fWnlffr,!l . N c utr<1phi b is(J I<tt cd fn1111 hL·parini ;.cd vc -
`IHHI ..., hh 111d I ru m hc;dth y vu lunl LT r\ hy <t nHH.I i fi L· ;tti ~ Hl o f <t prL· v iou sl y
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`o '.: (
`I , I!_,().: X. Oll : k:vc ls. I'M N denni es po lvnHlrpiHlilliCk <ll" kll k•'-
`CVIL'S. :f:/' < 11.111 v' co ntrol. ' /' < ll.OIII
`
`50.0
`
`Exh. 1032
`
`

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`
`RESULT S A ND D ISCU,,
`'
`· SS ION
`T hc prcsc nt stud y d c rn o
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`nstratcd th · t I
`ICt uccL th c kvc ls of ROS
`,
`l oxycyciinc significantl y
`<~
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`Cl 10 th h y nc ut ro11hil s and
`111 a cc ll-frcc systc m ·
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`l-120 , 0), we fo und tln t tl , .
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`<l ll ttto na ll y mhthit ed O H.
`In o st tox ,c ROs - . , . , .
`.
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`' · I hc Icsul ts oht;une d 111
`
`E/}('Cf of doxvcvcline o n 1he ~en er!/[ion
`
`I 7<)
`
`this stud y prob a bl y ind ica te tlwt d oxycycline do es no t inhihit
`th e ne ut rophil m etabo lism 10 pruduce ROS. h ut ra the r scav-
`e nges thc ROS ge ne ra te d. T hese findings see m to sugges t tha t
`doxycycline is cffccti ve in th c trea tm c nt o f acnc no t o nl y by
`inhibiting the growt h o f / '. acnes. hu t a lso hy act ing as a nti o x-
`ida nt s o n infilt ra tin g ne ut rophi ls as o hsc rvc d w ith minocyc li ne
`( ! ).
`We have rece n tl y re po rted ( I) tha t te tracycli ne . oxyte t-
`racyc lin e and minocyc li ne inhihi t th e Ieve l of ce rtain kin ds o f
`• 11 20 2 • O H · ). ge ne rated both hy neu trop hil s a nd in a
`ROS (0 2
`xa nthin e -xa nthin e ox id ase syste m . In co ntrast . our resu lts
`s howed th a t doxycyclin e rn lun :s th e Ievel of every kin d o f
`ROS ge ne ra ted hy bot h syste ms. T his see ms to suggest that
`doxycyclinc posscsscs the most po te nt a ntiox ida nt act io n
`within the te tracyclin e gro up . T he rcfmc . it is li ke ly that doxy-
`cycl ine is thc d rug o f cho icc in the trea tmc nt of int'lamm atmy
`acne .
`
`,
`
`R E FE RENCES
`I. Miyachi Y . Yos hioka /\. lllJ;IIllll ra S. Niwa Y . Ulcct o f <lll tihiotics
`0 11 th e gc: ne ration o f rcactivc- oxyge n s p cc i ~...· s . .I
`ln vc st l ) ~...· nnttt o l
`I <JW>: Xh: -1-1'!--l :'i~.
`/\kama tsu H . Kornura J . Miyac hi Y. Asada Y. Niwa Y Su pprc·s-
`sivc dfc<:ts of li llokic acid Oll nc utrophil oxygc 11 mctaho lism '"'d
`phagocytnsis. J I nvcst Dcnll<J tol I <J<JO ; 'i:'i; 27 1- 27-1 .
`.1. Sinico-D uric ux I, ( iougc ro t-Pocidalo MA. l'c riallin A. 1-Ltk im .1.
`E llcct of doxycycl inc oll oxygcn-dc pcndc ll t ki lli llg mcchallisms of
`hu m<Jil nc utrophils. Biochcm Pharnl <Jcol I'!X(>; .>S: IXOI - IX0-1 .
`-1. Niw;1 Y. Saka nc T . Shingu M. Ya nagitb I. Komura .1 . Miyachi Y.
`Nc utrophil -gc nc r<llcd acli vc oxygc ns in lin c'<lr I!!A hullous dc nll a-
`tnsis. /\rch Dc rmatol I'!X:'i: 12 1: 7.'l- 7X.
`-
`S. Root RK . Mc tca lf .I i\ 11_.0 2 rc• lc;Jsc from htl!ll <lll gr;ull!locv lcs
`d uring ph agocytusis . .I C lin ln vcsl 1'!72. (>0 : 12(>(>- 127'!.
`·
`(>. Klc hanoff SJ . Rose n 1-1 . tt hylc nc fonnat io n hy polv morplw nuclc<J r
`k ukocytcs . .I Exp l'vkd I'!7X ; I-IX: -1')0- :'iO~.
`7. Niw:1 Y. K<JS<lnla T , Miyac hi Y. K<II Hl h T. Nc utrophil chcnH>l<lxis.
`plwgocytosis ;tJHI paranlL't e rs of re;~c t i ve o xy!.!L' Il spL'cies in llllll\ ; tn
`aging: cross-scctio n;il anti longitudin al st udi~s. Li k Sei 1')')11 ; -1-1:
`1(>))- 1(>(>-1.
`
`Exh. 1032