The Unique Properties of Oipeptidyl·peptidase IV (DPP IV I CD26)
`and the Therapeutic Potential of OPP IV Inhibitors
`
`·,
`
`K. Augustyns", G. Ball, G. 'Jllonus&, A. Belyaev", X.M. Zhang", w.
`BoIsert", A.M. Lambeirl'. C. Dumxt'. F. Goo!!
`lib n A. HHme<s"
`
`d ~Q.,IIti'" _~allMdit:2lo!IioI:f$'liaIl'.
`D
`l.'! d~ /lAW. !.lot ... - :
`f. B-2lS10AoWa.padJ."u..
`Ab..._: n. .... _
`pl~_ ~a1~'"
`peptidas<I rv (DPP IV. EC U.14.5). lN5....-... ........t
`Ilan
`.. " ~"""genC020. n.. _
`Ul*U '"
`11M - .
`OPP 1V~D26 ... l» ~MC1 : Iho Slructur1l '" OPP IV WId ltoo .... lonWy
`~ belongs,lhe subslnlle 14l8didtY. Iho disl!tlu(cid:173)
`ol Mrin& p<OIeaMt 10 _
`1100 In'M "",","n tlOdy, ,,*,lfic DPP IV ~O<ll .nd ......... '" CD26 in 1M Intestinal Inc!
`",na1 handling ot ptoIIM conuoinlng pepIidM, in cell adhellon. in peptide metatlOlism, in IhI
`I........... "yotom and in HIV In!«tlon. Eopoor;>.Jty the l.I'I.1 "",,~nl' in 1M March lor new
`InhIbIIOB wit b& rtflO/ltd u _ .. t". discovery of new nan"", oubs!,.tes lor OPP IV _h I '
`1M gIuc.ol1O"".... ~_ Inc! thI <:/lfIm<>kIneS. r....l)o IIW ther~"" perspec1Wu l<>< OPP
`rvn-.. will»""".'"
`
`Inlroduo;lion
`
`D'; aPIA,. P"'P'i""M H lDf'P IV~ _
`alai
`01 ......1Il
`In eun-.nl ......fdI. Trw bJootoo.-· ..
`.. _Dill (.-l. 1 _
`1131 WId 1M ........,..". '" ....
`",'.:" in Ihe ~.~ _
`in
`.....,t :"11 (MC. 1n, 112 _
`113) .... _
`.....~
`. . doNI
`!he ,_..., OPP IV
`....A.d. _
`,,'•• __ po........,.... _ .......... 01
`.. "'lII' N,l
`; liM
`.... __ ... OPP IV. _
`peplklu and tt.. ~, _
`tt.. Iolal
`"""I_lIS coo""mitIg tt.. '* 01 C026 in
`
`~_HlV~_~
`
`lind Prollne·.pec:lIJc: Peptl(cid:173)
`
`Proline
`d ••e.
`0uII 10 "" uniq'" 11fUC1u.. 01 p..... am::w>g lfl<I
`,mine loCido, lhe polpliOll bond Mfo.. '" alte' • ~cIine
`..Iidue III ,"'II""'Y resillont l<l ""01<down b)' commcn
`prot...... The..iQrfI. , I' net aurpriIIng nu,l ","","",
`enzvn- II'nicipol'
`in t.... c
`1I' cI wc/l _ .
`TheM pmlin.-opoc;tic ~ ilI'Y
`ill'c>Mant ......
`In 11141 'eguIeIiDn 01 l!>e
`01
`1Iy ecliYe
`~'ll' 11~ Aema<UIlIy, _1lI_poo' M " " ,
`l!>e Jil»pflISK ~
`~ _
`~3..,l3.t)"",poall '(ECU,lUICdofa_
`
`""*"'·~ic .....>ymes __ <llIrncnI11lIlfMl.
`T__ lIIe Hole_ ..~H"_ p_
`~'iI_' 1'0' (EC 3.•• 11.~}. _,iii. r,,"I'" P (EC
`3 .•.11-'1, clijwplidrl P"ll'""M II (EC 3"'.1 • .2, DPP II)
`_
`IV (EC 3.•.11.5, OPP IVI. _
`" C·I"mlnal
`__'_1"......t>aoo,......·lu· P (EC 3.4.17.16}
`..... prcIrl .._ "..tl I' (EC 3.•.11.2}. The Cdof
`_
`. . . . .-11
`• ~ 01 t ....cllli"~ .. Pro-X
`, (EC 3.•.21.2&. POI Pl(cid:173)
`bond ilt prcIrl 7~ . . . '1
`n. -=* _wage poinII III _
`po
`...
`•
`- ..... felll-(t).
`
`I',~ect_t
`
`Struc:lure and Classification of DPP IV
`
`Oipepl~ .. fV (OPP IV) ilt II'Iig/'IIy $flIO<:f'oC
`",illCPtlltia_ dlaving 011 dipoij>IideI Tram !hi .......
`II"""",, cI II'l'lHei wilh Pf1Ile,."llIlI)o lII'cIine el lhe
`penuUiMlle poallion.
`11 Is e type II "''''''''''11''....
`ptO"I" hiving. cyI"!'IUmH;
`t.1I oT only 6 h~hly
`~onl, ...ed Imino .cido [3]. Th. hydrophObic
`t............-nb.'" "9iC" Is localed .tlhe N·t'"""",, (7.
`
`2'9]. ,. n.>.itlIIlla1< IinkIlIIe ......,.q_ ."'*""" wiIh ..
`
`lat9' lI'\'COIyIoollod ~ l.&-~.) .• qoo~ roogicn
`(325-562) and the C·lefn'W\II _In C<lI'UIninll1ll4l
`eatllyIlc: lriId (S!ioJ.-166) I.]. The pwlllto:l 4ft2Yl"'l1o
`.,.,1til:II __
`IounllID be - . CCfT1Jfiaing _
`01 l1Qol301IDll, - . .
`
`- ..............
`
`A .........- IcmI III OPP fV/CD2111 1_ .. II>e
`.... NgicnI_In __ 1 '.-"
`_
`0 1 , ._
`- . The po
`Icrm 01 DPP IV __ '......." ThiI..."...-.II,.,..
`0
`"-...
`
`.... in """'*'
`
`SAXA-DEF-00199
`
`AstraZeneca Exhibit 2007
`Mylan v. AstraZeneca
`IPR2015-01340
`
`Page 1 of 17
`
`

`
`_..
`(cid:173) ~i. I
`
`I
`
`•
`
`'" c........_CI>omIoO)', 1m I'd. (No. <
`
`~
`
`e•
`.~
`
`•~ •
`~ •
`
`~
`
`•
`
`~
`
`~
`
`-
`
`0i0a0Ild\0 _
`
`...... 'v
`
`"""" '.;
`
`COot>o<,F'I"
`__,,;
`
`"
`
`so
`
`~rea~down product of bour><l OPP IV. but sllows a
`signifICant .Irw!ural simlarity (5). """lilli' novel c.,~
`..,rfaee-.xpre••ed protein wilh OPP IV IIG1Ivil)' is also
`descrit.<!, and is ,marred to as OPP N·fJ [6J. R""""tIy.
`a memll,ane·bound enzyme with lI"la~na•• activity,
`seprase. was found wilh high homcIIDQY 10 OPP IV but
`with dille rene•• In subslrel. spO'Ciflclty [7]. The
`
`e"!>"",",,, of Ih.. prOl...... COfffl~l'" w~h 1"" .w......
`
`phenotype 01 human mel8nomo aM ",""'Onoma cells.
`A",,'her ,,,,,.nlly diocovere<l hOmologous proleln il
`Ihe lib,oblaSl-acti,ation protein lFAP).... lecliv81~
`eXjlftls.ed by reactive stromal libroblul:S on epilhelilll
`cane.", and he.,jng woon<ls 181.
`
`k"'mliliclltion ol!he ac!iv1I oil.....0:1..... in '.' Of'? IV
`by olfinity labelling and sil.·directed mulagenesi.
`allOwed to conclude lhal the ll<tiv1l .~. Mrino if; 1""'-"'<1
`in the
`sequence Gly·Trp·Ser·Tyr·Gly, which
`C<l<,eSl'Ollds to the acti•• ,fie motit GIy-X-Se,-X-GIy
`proposed lor ...ri"" p"'t....... [9], HoweY.'. DPP IV
`_
`• not exhibll sequence .Imilamy wilh any 01 lhe
`member. 01 the classical .a~"" protaa.. lamille',
`chymo1fyp.ln and .ublili,in. Thera 10 circum.t.nlial
`evidence thoat DPP III is a membo, of a new family 01
`..rlna prot...... the prolyl oIiooPaPtkIasa lamlly 11PJ.
`AJso pro!y1 oligopeplldasa (PO) and acylamlnoacy'r
`peptid.... belong 10 tN. farrify. shamg. """'Nsd 0(cid:173)
`terminal SI..,loh 01 app,oxlmataly 200 amino acid.
`oonlalning tha catalylic Iliad, The orda' or the catalytic
`triad _idueo i$ dilferell1 in the th," lamilie. : His·A$p(cid:173)
`Sar in ohym:>1ryp.'n. A.p.HIS·Sa, in .ublilis1n .,.;l Sar·
`A$p-HIS in rn<>IJS<I DPP IV [1 1), A oitrilar calalyllc Iliad
`wa. loond In lhe aifJ hydrola.a. which '.... roup .al)'
`dwa.... enLYme.
`',,"h SS lipsse,. choline.lera
`.
`dlenalactone hydrol...... dehak>gen.se. and
`,ina
`carbox\'PflpUda.a. [10.12]. Mo,,,,,,,,,,. Ihe predloled
`_ I ) ' struelural organisation ol DPP IV and Iha X(cid:173)
`ray sl"",ture 01 porcine muscle PO (13). ax""biting a C·
`
`larminal enl)'ll'lSllo <lornain ma<le up of atta,native a(cid:173)
`helix and /l-s!>Ml segment•• Ia .tril<Jngly aimilar to !hat
`pmvkled by tha th<ee-<!in'le<'l4ion1 'Iruct"",,1 sr,alysla 01
`alfJ hydrola ... /1(.17]. Llhwlaa,
`the
`three(cid:173)
`dimensional structure of prolina ....nopep!ida.. ,hows
`two domain., with Ih. larger <lorna,n conlalning thtI
`catalylic lriad '00 .howing tha glNIlI,allopo1og)o of thtI
`alfJ hydrolase loid 117].
`
`Substrate Specificity
`
`Tha cat'lylic rnecllanilm 01 a ••rintI prol..se Ia
`<Iepicled in Fig. (2). Tho> thr"" amino acids invoMld In
`the cstalyllc 'aacllon .'" ...rlna. hi.tiel'na and so.partic
`acid, They form lh. oalalyllc triad In which lhe
`nuclaophillo propaMIa. of tha ...rina .~ Inc.....
`d,a.tlcail)'. SUbstrata binding~, a conlormaliooal
`chonga in thtI .nlyrr'lll lhol leads 10 a prolon shih 01
`.arin. 10 hl.Udina. Th. form&d hl'lidinium Ion i,
`.Iabilised by Iha "_n"l. moioll)r. Tho ac!walsd _
`attacks lha cartXlllyl groop oItha scis.11e amklo bond 01
`lhe subsl"'la to glv. s 11,"1 talrahedlSl Inl'rmedlala.
`The ""9alMlly ""afOOd lelrahed",1 adducIia 'labili&ed
`by a hydrogen t>oncllng Iyslem In the oxy.nion _
`.
`Dacom,x,sWon 01 this inlarmedl.la gives an acylatad
`anl\'ma, whicI1ls ..,bsequantly hydrolysed by _ta, via
`a saoond latrahedral intarmedlata 10 thtI csrboxy'lc acid
`pnx!lIct and the SClwa aJllyma,
`
`TlIa sub.trata lpac~icily 01 DPP IV he. - . , ....1
`oharactarl..d, Basid.. proline .1 lhe penultlmata
`po,~lon (F', position) ~ can accommodata, alt~h
`la..
`afllclanlly.
`Ilanlna. dahydroprolln.
`and
`hydro.yprollne [181- In .n a.t"nclild InvastiO"llon 01
`the Sutlttrate spacdlolty using AIa·X·(·nIIroanllk!ml, ~
`was concluded thol lhe anl\'lTlll can hydroly.. some
`substralas In which the ring ",,,"lure or proline In
`
`SAXA-DEF-00200
`
`Page 2 of 17
`
`

`
`","_.01_01oo_..._. * -
`~C;:" -
`f"'
`
`--~
`
`0)
`
`-"
`
`--~
`
`f"' -
`r
`R-If0
`
`.,~
`
`.... 011 _ , '0
`
`_
`
`""12). Co<oIl"Io _
`IIO'IIbn P,'* bMn no:lild 1nlIl. qdIc ~ _
`
`~_ Tl'Os..-.ow. '*. '"-.., .....
`
`"""'.... 8QId by • high CItIIY* ............ On . .
`""'*"Y, __ o:4Midii..z.-.,k _
`' -
`1MriIy.tu
`_
`"- ........ ~in
`__ .......-
`S<lbalitulinQ _ """",,,.
`,roup by ouIphur or n"Y'll'" ., POIltion • 01 !hi
`VI" . ' ....... do"IlIy~.... ~ 1M
`efflnlly. '-e>...... llighlly <llIctHIed tor I
`0 ..
`....1ooIoI.ncl incrNH<l for .... lhla ....lI:'IlUt'. ('9].
`TltI "lHlm".... "ep lor ItII h)'drolyt/l 01 tut>"'etK
`wIIh Pro In P, 151M <I&ac)o\lIlloe, "'" lor ~1II In P, 151lta
`ecyIatlon rMCllon ['8].
`
`St..aiN with SliM Syrlthellc pepIIdn show 1M' !he
`P', ....1lIon~ II """'" "*II. e~ MCOMaty
`
`'""*'" sut:h .. ~lIod'"*"'..... ~....,
`~.... Anr l4tl'ino add (.Oft be plIcOfle'lIte
`P, poI:IiDn, .. _1Ital1l>l N-1ImlinII1nino k.nclian
`.. ' - _ probtIl". In _
`........ ""... 01 Ilta
`01 !hi P, "'*'0 KId "'" otIIy . . . (cid:173)
`...,. _
`!hi P, "'*'" aDd _
`...........~ WIlh P<oli'tI .. lite P,....., __
`.. l_"; _ ._
`"Ii." ~ ['Il- TIte CO"'" . ,__ ..
`.......... P, • ""'"""'""" 0 . - _ " . . N(cid:173)
`p8jllidIe ....... bio'w=, P, _
`P, ,.. II:> till __ tor
`\k-,/2OI.OPPIV ...
`_
`
`..
`'11.,l.2~__ ~'c1"..z~
`KId or ." n,.. or 11III
`01 prtlIM. 1M
`,._ 01 . . proIrw""'" by (~2-
`'....
`
`_l'k_......
`
`.. t dii."lI:>iIt
`
`__ ..-... ' . lolIl II, MI.' ~
`
`f"'
`
`~r--
`
`r ~--~
`A--
`r
`R-rOH
`/"--
`
`f"'
`
`..._~-
`
`.lhioOf'.... II'IIIID ....... tIIIIlO=, P, _
`e ,oo.IOOD loti-"*' ~121~
`
`P,. twll wilt
`
`,II..
`
`inlor"",1on ,""
`....
`81H<l
`p<otlel>'-
`1n ... e=..
`·,,0I
`......."
`silt oIllPP IV ,.. bHn ..-.:lI22l_ • mDdIl 01
`was propoHd
`!he KIM _
`_
`"I . . .., __ ,
`r.,,[ZJI.A
`
`_-....... -.....- -
`
`by 09P IV .el . . . 1>_'''. 1I,.oIwll." trw
`1nI....-,. by . .
`It''I' 'lion 01 !he lIIt1IhId
`1ormM..... of ..
`willi ... p...p, ...-
`bond "'"~ of . .~ _'V" by e
`proton ,renll" from In. POII'I•• ly ell",."
`...-.ote........ [24[.
`
`O[alrlbullon
`
`DP'P IV Is ._11<I ubIquilOUlty In rnammaliart
`~I ."" 0tgInI. II II tOUl'll;l .. ItII I<i<IMr 0:0",,,,
`~ ~ h..-. 11... 0 " at ........ ~.
`pane..... 1IIbrrIui"" gland end ~ II II
`••__ in I'lIg/I """"" In tI'IWrnbf.... re;ioftI
`0<0'''''''' u microIr.a _
`.. IhlI bn.oI/'l_
`......,. eo...-(cid:173)
`rwgoona III lite _
`• I
`.• _
`ofOPPIV..-,-in......., • • • ,.
`I I " " "
`ICIMty .... Iti;hlIII .. __ .......... low in
`I" ...... _1251- '"" 0IlI'rt
`tIuI nat I.
`_
`... ~II**-,II... -..,.._I
`-
`1I_~"'IhlI"""'_II_1n
`bo\IIIl bo",", ".'0._ [28J.
`
`SAXA-DEF-00201
`
`Page 3 of 17
`
`

`
`". _ _~. ......"'No-.
`The ....,... hQ _
`rwpor1ed in body ,..... ...."
`'JflOWiaI lIuiol.
`~ "'_1uMn tlOdr ..... onIy .... OPP rv
`.. _n ......... ulMl. urine _
`KMy ...., -.. Iound in ce_pInaI _ .
`'." __ P!"'I"... ,.,i,,",,_liIlu«l
`PI
`,.
`
`"'
`
`..... ~ iii._lIil Qdy on
`
`".,b••_oro Mi'll rr...,in_ .......
`-"-r"oPPN~.-y~
`:..,....,.0P9N __ Of.
`.........: "'"~C02e._ ........
`,m
`...."" --.gJy on "" II' ._ . _ ~_.
`... T<eII~......
`_F".........._
`on . . - ... bolI'l .cMl«l C~
`CD2e
`.m.... _
`_COlk T cell andCD2e Il:q)i
`willi"" CIjIIdy ... 111I T cell 10 ~ inIitrIlIukln-2
`(27). 0IIIliIId analysis oll<bM\s ol CQ.l. '"''' ...."''fIeI
`indIcat" lI\iIl CO;?6llppN-11lO De more rasttlcled than
`iTiOIl aeatUOfY _
`(CO.5RO.C029.)
`I~' CO~ m.molyl~.lplt
`pcI9Ul-'iO". T~i. un;q", popul-'Ion OJ ~uman CQ.l.
`cellllillhI ony one thai can ..opond 10 ,_I anl~.
`lncluc. 8"",,11 ImmllllClgloblll1n (lgGl Iynl....lif, InC
`_.~ MHC·raslr101ed oyIOI<IlIi: T _
`[2e.2Il). ""'(cid:173)
`'K....Iy. _-densily ....... ot C02e 1'\01.... _
`<k1_ on 01 _~ cella. ~1Iy
`_
`I:lO). 8 "'........,... [311 an:! myeloid
`
`opp IV Inhibilors
`
`-""
`0P9 rv illi._ by'" 'I'PQl_ po , _
`.1_ 10 u.. prok-4""""" In p •• causing ........
`1~Dl'ly po_ (Fog. (3»). ThlI II noIlIlIJII\Sing stnc.
`~ I, w.U k........n t~al I"'plidfl wllh Prolin. I. II-.
`lo,m ,1I.1I..ly
`...y
`p.nuUimll.
`pOlillon
`ot t'" N:tlrmin.1 n~t"llln on I
`dikIlOPil>l.'zinos••riling lrom t ~ allaCk
`.....,. c.rt>onyl
`
`ol"_, lor_po ,
`-...0py4 .......... 4 ( ' " /9l. ... __ -'ely
`p3I. _11'IOII'" -.... pepli:M InIIogo.-
`, ....... _
`...,
`.. ~ ~.... P,~_ ,..,...
`. '" 111I_ ol OPP
`IIW '101"
`IV.' cydizalion I'UC1ion can
`..n 111I " -
`amino lliOUP Cl1III p • ..-..o __ 1III_rq>/IiII
`
`-
`
`_
`
`Xx·,
`-,
`
`OI'P N i.,·
`III _
`fls. PI. _ ' I i
`li
`' __III"" . . . ..--..
`
`• _ ..
`
`--(cid:173)
`
`[3C~
`
`ttoo IhInl ....... _
`CI "" ""'''!c'' .....
`...,-_•• lot DPP IV iii_I.
`OPP rv ~._. _ --.. in Fig.
`
`De_ '''' M<»tlIl _
`n
`f<>. only • _
`li.·'.,:
`po ,
`Typical
`(el·
`
`IIW.....,... . . . .onIV_. ,:g, _OPP IV
`.. . . . . . ,_ ... ~. ':g •
`-
`. . r.yo"""'ldionI ol
`~ ('J..uIing _
`inhlIiIofI. i_, c:cnslI"" ... _ .....,.-.
`pro:lua i_ i III in . . rIrogI CI K,. ,(l:2OClO I'M
`(IlI,PJO<U•• 2) II. ~~... inhitllIot_.I<j. 2.2
`[1.23!. Ollgopeptld. . .1111 N,I.,iTllfIIl
`j(,p,o
`HqUenl:eI . . . .110 ""","1irnII inrIibIlora. 0ipr<1Iin A
`is "till, 'U'P"'~ .1"". tl-.
`
`Pro<lUC:I,IiU c0""4'OU'ld'
`
`lI<:klng ll1e carOOny!
`
`I'M (351, T~il .., ....
`C<ltnpOUrKl ~.s t~e ove,aU SUllllrlll,llka llruotu ..
`M_ lo< DPP IV hyd'OIySI•. A ""'" ""'n! r'POrI
`er.ow.o It'll! tl>oH tripeplidl.... if>dH(I .ullsl..l..
`with e lOw klrntl¥Ir notIlk-) [31'11.
`"-"":lion 01 111I ""*'" ..-.. IUC!l .. a",lnOK"
`prm>!lcll......d tlli&tolldlne. __ ~ ...
`DPP rv inIIibiIofI [23.".315~ The
`IJola'Il an:! _
`_I POlen! ~ ... WI __
`N·
`............ ~.In._~
`~ PI-"1Cso __ 2.8 I'M.
`(~00If~...,.5_1
`........ NI .. """ •. -.. (.)_
`N-(~
`I0 ......_-.go_..
`iI In.oe too tile ""'*
`.....,.dIcfIu«l"
`pooDon"''' ",,,.-. ring
`
`i&flop it..
`"
`·Q"0iM ~._
`. - . . 11>1I cIII"I"'Q ttoo 5,
`_
`ling ...
`- . . - . '" __ 11-.... 1 ,o•• boi«l
`"'••_
`-.go
`os
`PO!I'/'ICr f301. TI-.
`II'IIlOgu<tI. Introduction
`
`IUtlIl~...nt .t till 3-
`
`,.,.... IUlloIlilu1nl OUCh 1$
`_tit)' 1CI!olly. Only •
`1IuorI.... II 1_ 171. To ""b1illl In optin'IIl N,
`t~ reIltIo.4....... 01 "'*"<>IcyI ", ••_
`.. --.
`P"PI'Id.•lIowlng tllal ~P<>PhI~ .mlno 1Cid0 ll"....
`more potenl oompound. [.wI. In pan""ia,. /l-Of.ncllld
`o·.mlno .cid derivative. wI.' I~I mool poten!
`oompouncII wit~ till non·prOlllnogenic Imlno Kid.
`cycIoIIe~providing till m<III acIM ",,,oIididoo
`(I).
`
`.erie.
`
`Ipplled '0 I
`Tile.. linding
`[<10•• 11. The~
`pyrrolld_2-nilri
`'*blllip lot . . NIa"..... reIltIo.4 dl I,'" in . .
`..
`PJiii"""'" _ ~«l wei lor
`- . _
`tile "'C4l polerI c:ornpounll bWog ....
`
`K,. 1.1nt.l_..
`~~eil.. _lioI t. lltII~ '*.
`I.-,11II pH.1,. It....
`
`• -.
`"II).In--oto __ ",,,.,
`ttoo PJIi ..' ....,2,nIt... to ""
`, ~1-- .....t11 0'
`
`SAXA-DEF-00202
`
`Page 4 of 17
`
`

`
`-........."--_..."""--
`
`,
`
`,-/-"
`
`1
`<
`I
`I
`7
`
`v.s
`y.OI,
`'1'IfZINOtl\. •• S
`,Y'IZlNO,l\. .. CH,
`V. <:>II'
`
`a_._._."01,
`
`•
`
`"
`11
`
`, •• 0
`
`v.""
`
`"
`
`"
`
`Fig. (4), _led DPP IV inhibit....
`[42].
`tMj..olid;~e·4·n;Uile e~h.nced pOleocy
`Replace"",nl 01 the amide bond wilh a~ looola,oc
`alkene in lhe l$Oklueyl analogoe (10) dec,eased
`polenq a 1000 fold (K,. '.7 11M) 143).
`
`The HIV·1 Tal (I-SS} ptateln has bH~ rej>O(t.cl
`a OPP IV I~hlbjlor.
`aoggaatlng lhal
`Iha
`as
`Immonosoppraillva ellaclo 01 Tal O~ no~·H(V-',
`Inlach"d T .,... coukl be "-t.cl by OPP IV «.4.-45), ~
`
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`Page 5 of 17
`
`

`
`". --~.,.............
`wu ._n INlt n.. N·I.,mln.1
`I ..·I..·P...
`""**".....~_.. _B
`"• •_ 01_ pn>Min _ ~ 1M1"...,.
`_
`
`in ..... _0:00''''-'.' 0...... 1-'8..7).
`
`_..." ... _.~~-,...-
`
`laa-bo<oPro dilMpU.... ('1) __ ~
`in !he
`MIOIIll!he mosl poIOnI 0o'llbiIDi I wiIrI ... _
`
`•ranaiIion Ilale ~ wiIIl slow.
`IjgM-fIirding
`Io:ir-..rb jqJ. The ~ P-ottIiIIl • IloIon '-1Io.IghI: '"
`....
`10 Iorm • _
`~ _
`"'" calatylic _
`mimic:I the IfIIntIIIon 'lOlO' 01 omidoI
`compIO. _
`"Y'drc>lytit. 5epo",1on 01 L·Pro-llt...-ro Into ~.
`dio".-,. sIIowed!hol "'" L.... 1tomo< NIl. ~
`v..... 01 16 pM ['9~ Unl<>l1UIII1'~' lhoy III"". tI>o<l
`h.W·lif••, ...... "1 PH. cauN<l by cyclUllon of lho
`t.rmInoI .rnjno1unc1ion wlt!1 tho boronIo ocid. f<mring'.
`eydie. Nc:tMI speciIt C>I:>I1lolni'lg. 8-N _
`(F;g. (3}1
`150-52). IrI • SlfUCluro-ocIlvity ....'io1'o1Np 01 boronic
`v.1IeIy ell-
`ooIcl inI'liI:>il<:>f
`-"!hot • _
`........ _
`I<'C1fI*d 11 !III p. ~ but 0.
`........ tcidoI. "'''''''''''1''- an*"> _
`..... glycinII
`1_.led /531. no. '~lClfd, 01 n....
`.ro _
`•
`~IorDPPrvo~n,. ,_iii
`- .""'"
`mDfl~ __ ""on
`•
`lor PO u_, __ :JO III
`DPP rv __ oIso poIOnI ~tbM: • 01 0Pf' I. On 1110
`0:00",.,1110 ICso _
`IOOOk*l ,..-.
`
`---
`
`nell t..
`th4i.. co_ncb
`"'" IIIgh _
`01.
`. . . . . - gc'"'ld Mo'llbiIDil ISS!. Sorno
`...... ol ..'..._.-.~.....
`.i\iI*.,.IJ4r_dooo- 1Ii,.. oIl,••nPla_t.lIo
`_ _..... ..-.g i1"'_""
`""MIr in lho
`ii.' ... tongIli /561.
`Gio.'ll. C.""""'-
`s,nIt .1": Pi'!
`)'1..".00: ooIcl . . . . _
`o/c:oI'IoI . - - '" P "
`........... been bJnd '" be POlenI ~
`- . . " . 01 o;hymoIryptin .na ......... The..
`M '*llf.ln -.., *""'r. -...-.,
`l' _...
`'II
`• ""'g'~ p_
`'low "Y<!rotpl.
`_
`10
`01
`~_--... 1 !e[$TJ.H¢ "".
`principIo .ro noI
`~ <lInigI.., _"ong _
`'o'I<y ICIiw II OPP rv oH:Nloo. (12 : ICao ~ 2.' ...... '3:
`K1o.1.7 .....,~ ...ub).
`
`""_wi group 01 Of
`
`N·pepUlI,1 - o· ,••nll.oblnloyllllyll,oxyl._
`",fn. . .,. .nlynM.eelivoted Wllb/lo,. 1", Mrino
`~ 1$8~ AIIor .....1Id; oIlh4i ~ ..........
`tho Inhfllil'" t""", e lO'l.nl chemielily , . ._
`i .." ."Me._.ItIimI.
`e
`It.ot.d'ng
`'" ...~ ... . . - on2yAW. ho PIa 1110-(-4(cid:173)
`~(l.li5_ ... . . . . . . -_o l
`DPP rv ISUO,. 5mIor~ _
`... ~
`iIIIy aft hill '" . .~ ooIcl 01 . . _
`b
`.
`CfaIin 01 . . p. MWIO ooIcl 01 .,.-=11 ...... &5_
`(lS) ore'" 0,.' •• 1_. but 0:00,'-' ~
`••_ , - K, _
`..... ,.....,_ tongIli (lil~
`
`Acyl .....,.... ~."'e , ~ ... torrn.tion 01 on
`.-,. 1hon 1110
`IiCyI .....,.... """'" deocy\I1ft _
`patWIl .....,.....pn><IuCl compIO>;... 4upept_ aro
`.n a-corbon '-~ lor • No
`ptlllide. in _
`• 10/11.••Nilrop/lonylofl.,. 01 fIlCh poplid.. or.
`In/libilor1. eI _,oJ ..rille prol__ III~
`ot
`on..,.... [SoIl. Gtycyl..........nyl·.~.t.,.
`lho lCli"" .it. serine lormlng • 'l'bII acylaled
`...ro!O\.lf"<l to acyIal. opp IV. but tho ""acylalloo ,ate'
`
`DI ptjd_.....~d dlp_,' p/>o,p"o...t~
`ul
`ore Pot*'l~ 10. '1"'< .01 opp rv
`1S2~ JlfObeoOfY tl6co.... tIw)' Iud 10 • ~
`....... 11 ....... Ili!e tF"'\I. ($)). __ ~.1Ibilot1
`onI'f • "oction (10 ""l eI lho .....,.... IC1Mly' _ •
`,.Initcl .Iler • WMll5 'II 0'11.... InOk:Ioring !III Itrong
`~ lnhibilion (63), In. 11l1Cl\t on !hi roll 01 !hi
`f'l .mino ackl In llipoptKll lliPh4inl'l JlAOIPnolllte.
`[63.&fI. K.... lnown lhot pmline In lfIII p(»lI"" ~
`
`I"-+--0
`
`•
`
`",,,,••._,__,,,••=m__.'·.o""'''i1,.,. '. _oIOPPrv_ ....... _ : ...
`
`SAXA-DEF-00204
`
`Page 6 of 17
`
`

`
`at
`_
`iftIIlbllO" (111. A .......
`the mosl polan!
`- - . ot '1_ " , •• """""" IltalIRt In luran
`cit_ pIMmo (1,/2 • S "I _"'" III ... _
`........ '" dIoi••N.... '"" ItabIify ot,. in pIBma
`~ dislM, ...u:.d .1Ii*" in ~
`..... b ...- , . in_... - - . IM_
`cor',*""IO~.n.._ .......... ....,'"
`a Poo-Pro •
`10 prol~
`b<
`' ......
`.... II: III.in-,
`trJ' ",d'.1)1iI of
`01 ,. II
`in" cause<!
`the
`.. '11 ,
`• '" a aingIa
`,.
`~te
`I 0 """ in ..tItIlII rtduca Iha
`_
`01 II (I. S Of
`...... OPf'IV.-,_ ......... IO .... _~_
`IllOfell'lan 20 dol'S !Of compIal••.co,.,. No! onIt
`
`l>u1 ..... DPP IV in
`poll..... 0f>P IV _. inl\itIiIed.
`cItwl.Itil'lg l'fmIlho<:ylM and perip/ll'al lit"," [65~
`"The role 01 lhe F', .mino aelcl w...... _'i9ala<!
`IIIowing IhlIt. 6-mernbered 'i\liIOgUti (i'lomopfOlinfl)
`lne....ed .ctlvity 162]. wh.,..1 .llnIne <ltC'U$a<!
`iX>tl<'Cy 1631. Wilt11... Iud COflW<incl. we ra<:<IfI!I'1
`"a<! . . .lies 01 diAry! '-{S)·proIy1pyrrolidino-2·
`IIyrt
`(R.S)~laI with~""" IUbItauenll .... ,""
`atyl tingI: ("""".... -.:r.I<'jWmo.~.
`......... "'.IIIO"1C"..t>on.... ancl
`.U.yt.minocalbon....)
`(~ 1. A good COfNlIIIion _. Ioo,nj
`beI'w.." ..... _
`__ 01 !he~
`~_, ~Iftd1lItIIitr. ThiI..-._
`and
`_
`lIa.Mug ....1.. _
`........ polInqI.
`"The _ I "<Wng
`bul also
`stabilil)o.
`....geo .... ot
`"". I
`. ,_ " , high jllllIfq'
`"".G1,,1CI wiIh • high u-, 0I1ha 4.-,..rn;no
`• coa...__Ilw (11.1C1a. 0.41l101. t"". 320
`min). ThiI ~;o apaeifitb DPP IV._1ow'
`c'(lOlOxidl, In I'unan ...Ipl_ bloocI _
`eel...., __ po-," '" ofwo. The""a.
`wa
`conlld" di(4.a.".Mldophan,l)
`1-(SI(cid:173)
`
`~t-(R,~(I7)..a"""
`Iurlhat11_...."'" biological 'l.O'ICllon 01 "'" ....,.,...
`
`lmpoo""iI" _ . PIighIf"- OPP IV innibiIo< ""
`
`IMibition.
`ItI
`01
`tn.,.pau,ic valu.
`.nd the
`Pho.phln.'.. smla,'o lhaia phoapIIonat•• we..
`• 110 praflIracl.l>u1 we,. poor lnIIlbilora 01 OPP IV 11.:
`K:1lO • 2.4 mM. unpobIlshad .......l,
`
`Anolher ._h to inac1"'all l1li ""rma • 'hi
`u.. ot Jollch..1 .dditlon .ub",,'''. The..
`oWh "'" ...... lila
`COI\'lp<UlCII could poIlflllaIy _
`Mrirla viii • Michael Idlitlon ralCllon. On Iha _
`I I an _
`--..
`t>on<l can _
`_
`• Iha _
`" •• If: A ..... was -rar. but _
`COIf"ClCIUrIlIs
`b.I>hlCl .. ""1P01i6.. ~ In Iha -...a,
`""llI. n. moaI polanI"',; : n:I _
`... -..",
`.....- a am.-
`"1 ._ (11. ICs.o • !lO jtM), _
`"".,...."""""""a rapid aqo
`doIpao_. IMlbiIion .nd _
`0I'IIy par1ia.,.
`bill . , " " " ' __, __"""'''''*'
`II __
`
`.ICI _ - . ~ ,jng tllaing Iha
`be _
`............._~ J.
`
`__.:-,.. r_........... >1,
`
`Cl ....\lOflIIIlJ'I~ _
`.. aIlrilr labaIio",
`.~. of _
`The,- .re IUbItraW
`prola
`...,..;,.a ..coat
`.....
`•
`I:;
`"
`·
`"'
`inIil!he adiN sill. Thwe. _. . - wilt!
`whdI
`_ of",. = .. eata/jtic _
`ot ... ....,.,... _
`-.-..", bklcU onzyma acIMIy Olp.plld.
`~ _
`.. __ .,........r 10 __ f66J-
`bJ' ....... I . .
`_~..... I "'
`cy:f
`.1Ia. dlpepllct'(l _ i l l... _
`I '1
`
`(211) b .1_, 01 OPP IV
`.....,,)'l blon.. ateill_. in Iha moc_.
`
`py<idinkMft
`1'IlIIl.-.
`
`""'" .... _-bncIing ...........'" _
`......., ,0
`1I7.1;8J.
`RKeo ....., • . - r,pa 01 n-rPJIa cyclopsp!id.
`Inhibitors w.,
`reponed. con"lning •
`IIl.nl
`QUlnouimiu'um mathida aI6cI""*'la (i') rflPOf'iIibIa
`to< inI':,ivalion (69]. This COf\1POUI'ld hal • K, • 85 nM.
`and. "",me!<If OF'F' IV. giving. '.pid Ir"va,.it>Ia
`irf'Obition 1/'Ilit 11.11,.. _,al "",,,•. Reh'llir1<ab/'f, OPP
`IV.~ .. much leu "NICted. con!lrming tha, ~ ...
`dd'1fIN ptOIIin. '6 Os an aruyma_aled _ " ' .
`';ne. anack of
`.eri"" at
`the carbon,1 01 ,'"
`unmask aI"', ...... hiving a good
`c,de~lda_
`
`'*'* ..WIg 9'0<4'. F<lIuwW", a lao, eliminalion of
`
`d""""Y'" fik tI>ia __ gl... a QUlr..........'"
`ion, _ ad in !he ao:tiva' .... during lila
`_
`01 ...~ trJ' __ of "'" papli1a
`_
`_A"''''lJpJ-.oIa-.t", I :;'.
`1M "" IIIiI 010. IItad
`in Iha - . '" ",. _
`II:' 09l'0iIa _
`..,
`.- h • __ ~
`_ ... do ... WIiliI ...
`......... - . "The __
`lall tIilluIlon of lI'lI
`.....,..... probably bICa..- •
`Il:b09l'oila _
`01 ""'..- _
`In
`,
`po
`,
`""""""" . . q<:ic _
`trJ' cydUIion _
`to _
`01 Iha 'rae ...-
`...
`
`-Inleallnal
`
`and Renal Handling 01
`Proline ContainIng Peplldea
`
`T'" .tlIO<ption 0' recycling o! Prot"'" c....t.lnlng
`peptldel. a vit.1 ptIIOltM. OF'F' IV'.eont,_ 10 IhiI
`procus • twoloIcI due '0 Its bn.oIII botOIo locaJJaaIion in
`l1li """. lnlulina and "'" kidney {vida - . ) . II pilI'S
`an oIllIgatory .... in l1li __ 01 paptIdu in Iha
`\)InIfating !he
`.......n.
`Inlfltinll .nd renal tubuli,
`1UbIltI... lor "'" paptida I;lICiIic ,ranapon •.,.._
`geo....... del' 'I • in OPP
`[7OI.lblng rail
`_
`IV. Iha IUU.liooill
`01 ... .....,.... in ..... t.........
`1"=. 'o"'.'('-5)(T}",
`Iftd~"'
`~in_~.....ib• •
`\iIIicIaa _ . n. "••ib• • ¥MidaI _
`!:!•• rat. .... _
`OI'P IV
`10 ",dlolj. Illa
`pentapa9lid<t 10 di· and 1I~ will> the
`lranIpOrt 01 _ ~ via ...
`. . . . . . . . _
`H- Il'........,..".._. b.......18i. _
`. .
`
`SAXA-DEF-00205
`
`Page 7 of 17
`
`

`
`_..
`
`and tripept>das. buI ""t tetra~ or ionIIer. On ttla
`con1rary, OPP IV.~t'" rats fallfld 10 hydrolyse Iha
`pentapepticH and he"", lackll<! ttlo abiMy 10 trans.po~
`the radlolabeled pepli<!<l [71). Funharmore. urina
`analys;' """,ale<! an increased excr8lion 01 proline Bfld
`hydroxyproline in OP? IV-deficienl rat•. Also. 10110wifIQ
`intra.e"""s admlnlSlration of Tyr.P,o-Ph<>-Pro·NH2• a
`peplide Ihat
`i. . .olusi.aly hydr<>l)'sed by DPP IV,
`urinary e.cret;a., 01 ttlo Inlact peplide was many-iold
`grealer in these DPP IV-~tivu rals.
`
`OPP IV in Iho sman inleStine has a similar fencban. In
`""'" jej..-.al perlusion of a ".,.,.,.,. tetrapeptOde. Leu-Pro(cid:173)
`G!)'-G!)'••I>owed thai
`lau-Pro and G!)'·G!)' ware trw.
`found in t"" lumen f72l(cid:173)
`major hydrolylic product.
`Experimenls using tha DPP IV_nagati" rat. also
`con1inned t"" role ot DPP IV in the hy<lrol)'.is ot pfOll'l
`pepUde5 and the as..mllalion of proline-rich proleillO.
`T""ra wa. no diHarenca in the growth rata botw...n
`DPP IV·nall"ti.e and comrol ralS fed on a relerance
`diet not rich in proline_ When t"" proleln sour<:e was
`chang.ed to gliadin. a proline·rich protein. tha con1rQ1
`ralS rnaintajned tna;r body _~hl. _",a. Iha OPP IV_
`detjdent (alS experienced a lign~icant weil/hlloos [73).
`
`Role in Cell Adhesion
`
`The abinly of DPP IV to interact w~h PfOleins 01 ttle
`eXlracelfular malri" as 8 cerr adhesion moH!culll he.
`t>aan amphasise<! ~y lhe basic _rvabon thel the
`DPP IV substrare G!)"Pro-AIa inlerleres in volm with lhe
`initial spreading 01 rat hepatcx:yleo on 0 motri.
`consisting 01 fibronactln and collagen [7~). Morae.or.
`an antib<>dy against DPP IV delayed !ibronectin·
`mediated adhesion of ral DPP IV·.hapatocytu on
`in .;"0
`denalured oollagen [75).
`II was reponed Ihal
`fibronectin binding of DPP IV was Independent of ~s
`""lOpept........ RClMty [76). Uk"""..., QPP rv an murine
`firlfObl4.t. ""18<1 as 8 coUagen recep1c>r [77]. On humaI1
`T cell•• OPP IV hn been demonstrated 10 be a
`funclional collagen r""eplor laading 10 cell acti';ation
`[78). These resullS show conelusloe!)' thaI DPP IV is
`able to bind collagen. and it has been raponf)d mor.
`recently lhallhe binding sUe is tound in the cysleine·
`rich rogioo 01 QPP IV [791.
`
`Due to Iha prasence 01 Gly·Pro sequenceo in
`COllagen and fhe higher secretion of proline· and
`hydroxyprohna-<;Qf1leining polpticHa in lIle urine 01 OPP
`it has been .ugge.ta<l that OPP IV
`IV-""llat""e rot.,
`plays. role In collagen melabolism. This gelalina...
`.Cli.ity would ba po•• ible If
`the anzyme has
`endopeptidase activity. ne<l to tha well eotablisi>ed
`a.opeptidasa ItCtfvKy. Indeed,
`It has been reponed
`.e,y recenlly lh.t
`ral DPP IV nhlbllS weak
`endopeptida.e acti.ily. a~la 10 cleavo denatured
`librillar collag.n•. but not nali•• "<>Ilall""" [SO). II hao
`
`the endo' and e.opeptlda...
`thai
`bean .ug518sted,
`activity reside In a common a<:live .ite. and thai
`endo!>eplida ... acI....rty should be """" in cantut with
`other gelatina.e. and o<>llagena ..... or ahemati"!)'.
`lhal this a<:llvity mighl he" au.iliary funCliono in call(cid:173)
`matn. ~s>on proce..... by urvnaslcing bir<ling .~""
`lor
`Ihe integrin receptors. The delectlon of
`Ihlo
`518lalinolylic activrty, makes h acceptable 10 propo...
`lhat DPP IV and the horTlOl<>gouo prolain•• ...."..... and
`FAPO'. reprasenl a naw .ubfamity 01 galoti""lylic
`inlegral membrane serine proteaseo [7,80).
`
`Role in Peptide Metabolism
`
`A lot oT cylokines, gro .... h lacTors and some
`neuropeptide. share an Xaa·Pro mollf at their amino
`Termlnu. [2). This sequence may nol only oonlrlbula 10
`l!\a biological activity. but can .Iso seMI "" ... slru::1ural
`profeclion against non·spec~ic proteolylic degradation.
`The slflldng deg",e 01 conservat;a., seams 10 refieCl an
`8Y<lMionary pressure Iowan! ttl;' Xu-Pro molh. On!)'
`lW<> proline_.pecific aminopeplidase•• ,e'pec1ivo!)'
`amioopeplidasa P and dipeptklyl peptOdase IV he.e.
`substrale specificrty low",ds lhi. Mtarrninal Xaa-Pro
`molh (Fig. (lU. and may theretor. be Impohanl in Iha
`modulation 01 the bloloQ'lc8l acti,,!)' 01 some of lhe
`CyIokineo, growth lactors and neurOllRptideo. Anolhar
`interesting oboervation was that p"osp"orylalion 01
`serina Of IIlraonine in a Sar·Pro or TIlr·Pro .ubstrato
`decreased the ~ value with more then lW<> orders
`01 magnltuda [811. This could Indicata lh.t
`posttran.latlonsl
`phosphorylation of peptide
`saquahCa$ may modulaTe ,na;r proteo/ylic stability. Wa
`wiU discus. hareaTter the bloIogical!)' a<:li•• peptides
`which ara proven 10 be hydrol)'s&d by DPP IV (Table
`(1)1-
`
`It was demonstraled that oligopeptldes with
`sequence. analogous 10 the Mlerrninal pan 01 humen
`interl.ukin_'11 (ll-111l.
`IL-2. lumor necrosil
`faClor·1I
`(TNF-JI) and murine IL~ were hydrC>i)'sed by purified
`DPP IV [821. In contrast 10 these ",suits no degradation
`was found lor the inlact recomblnanl cyIoklnes. Since
`Ihrea-<iimensional struCIur•• show that Iha N-termjnal
`part i. af the surtace 01 these proteins and hence
`st>ould be access.,la to exopeptlda.e •• additional
`steric factolS musl Influence thei, .usoaptibility 10 DPP
`
`•
`
`II was lhown lIlat puriliad OPP IV hydrol\'.a.!he
`nauropeplide "ubotanca P [83). The undecapeptlda
`
`SAXA-DEF-00206
`
`Page 8 of 17
`
`

`
`_.
`
`_ondN-T_,--
`
`_"'-___......--HH;/
`
`'''"'''''1''')
`~c
`T,......•..............
`
`....."'. i"
`T,......·~--'
`,.,......-.......---""'>GIy-
`""4'ro~___.
`--~
`__ ",""0." .-
`r. ... " •
`-. ., .....---
`l)o.-.__....n-.-._-
`-. • '__'(7-311I
`T,._............n.._._
`-. •-_.
`_.~n......n._.....,.
`-_.
`-_... , I.",,_·,~
`_____on_
`
`_
`
`...........""l
`
`1tI-.....~__.Aop--<)U-
`
`........__ n........
`
`1tI-....·-.oor-v_n.-_.....,.
`s......-r,.._._n-w.n.-._
`
`l........'V...~.....s..T,..~
`
`~
`
`""'"
`..
`,
`•
`•
`•
`,
`•
`•
`•
`•
`•
`•
`•
`
`......-
`""
`
`(11,"-
`
`".~
`
`-
`..
`
`""
`
`~
`
`(,01>,""1
`
`['01>'''''1
`
`".
`,'.
`
`I'O>.'O!II
`
`r...'IOI
`
`waa co,wenoo 10 the f,agment (3·11) ana (So") by the
`sequen!ial """",,,"I 01 Aro-Pro and Ly:s.Pro. In .<ldition
`to plum. metabolism 01 aut>stsn<:e P by angiotensin
`con....nlng enlY"'" (ACE). the majority 01 hy<j~s
`was due to DPP IV. In t"m. the wbslanol P (So11) was
`rapidly hyd,otysed by Ir.-Wnopepli<lase ... to subs'a""e
`P (6-11) (&4). Using OPf' IV-IlIIlI"live illS. ~ was o.hown
`the, .Is<> .., vM:>. OPP IV w•• ,,"sponsibll f<>r sub".""e
`P hydrolysis laS]. A comblnetion 01 an ACE·inhiblto,
`(e.ptoplil) .nd • OPP IV Inhibitor (diprotin A. 2)
`prevented to • Ia'ge e.lent substance P dsg,adalion In
`plasma [8-I.8SI.
`
`/I·ClSomo,phln (l·S). a peptide wilh potsnt
`moophlne·llke bio~1clI.1 activity dsrlved 100m milk
`proteins. Is rapldly hy<jroiysed by OPP IV to Tyr.P,o,
`PIIe·Pro ..... Gly. Thl' cleavage CO<JllIotely .boIished
`the activity (86). Changing Pro to D·Pro Imp""'ed the
`'IatNlity both ... vitro ..... .., vivo (86.a7). Df'P IV is a1..,
`
`involved In lI1e hydrolysis a.... mnal ..plake ot Ihi'
`opioid pepli<le 17'.68]. Together with ACE .nd
`ca!t>oxl'l"'ptidase p. OPP IV i' responsible tor the
`int&Stinlll <klgfedsrion ot jJ-casomorpl>in 11·5) [T3].
`
`Tile same SlrJdies repon tile Importanee ol ,....1
`OPP IV hy<jroiy$is k.. mo,pllleopUn (Ty,.Pro.PIIe.
`Pro.NH.l Also, In situ perfusion o_riments In Intact
`animaJs, revealed that1he s!;Iility 01 the opiale agonist
`rnoophicoplin, wilen .dminlttered 1n10 the Inlssti""t
`to block tho ehOlo'.
`tumen.
`,o.in·Ir>d"Ced w.te,
`seclll!ion. was slgnffic&ntly greals, In OPP IV·MllSli....
`",to (73].
`
`M."."... 01 the p.nc,ootlc polypeptldo I.mll~
`are slao s"bstr.tes fo, OPP tV. P",ifiad DPP IV
`liber.ted Tyr·pro trom both. n."ropeplldo Y .....
`pepllde YY l<> thai, (3-36) fr.gments. but .Imost ""
`Ala·Pro from panersa'ie polyptlptkle (a91. This was
`
`SAXA-DEF-00207
`
`Page 9 of 17
`
`

`
`contirmed in ...rum, and 1I'e hyQfOly$is was blocked by
`5!>8C~ic DPP IV lnt1ibit<m, Removal 01 this N,lermi<'1al
`dlpepliM ina<;livates the... p&p1idfts lor binding 10 one
`receptor subtype. and is therelore Impo~ant lor Ihe
`~I activit)'. !lfIsid8 OPP IV, aminopej>lOdase P
`and endopeplidue·24.1' are alsO involved in
`proc..si"ll 01 these ~tides 190-921- Peplidft YY (3·
`36). which has an N-te","nal Xaa·Xaa·Pro lO9quence
`has boon reponed as a competitive inhibitor 01 DPP IV
`146J, ~ oome kind o11_k r9gIIIallon,
`
`The degradation 01 anu,ollalln. a potent
`inhibitor 01 100<1 inlaka. by inlesllnal brush-bofder
`mermra".,. and brain mermranes involves both OPP
`IV and cart>ox\'PllllIlda.. P. In serum. dll'Qra<lotlon was
`mairlll' <!ue 10 DPP IV (931,
`
`using DPP f\I·negative rats. ~ was shown thllt GIP and
`GlP·I (7·36).re degr.ded by DP? III (101]. M...
`specttomelry Indicated Ih.1 Incubllion 01 these
`peptkles in serum "!'Sulled mainly ., cleavage by DPP
`IV, ....h onI\r minor HOOndary degradation _
`to <>Iho'
`",rum prote.... activities [1021. This kn<>wied{le Ie<! to
`thl _lOpment 01 GlP·1 lnalOgue. with a.<le_
`metabolic Itability and Improved blologicll activity
`1103J. Furthermore, ",al adminisl13lion 01 tho DPP IV
`Inhibilor 11e'lhlazolldlne (3) to Zuoker ralS ,""reased the
`orlbitdy 01 GIP and GlP·I (1041. A similar elllct wa.
`....n with .n InhiMor in .naeslheti.ed ~g. 1105).
`Gluo.gon·llke paplld..2 (GlP-2) is degraded and
`in .ivo. which led to Ihe
`lnaclivated by DPP III
`develOpmolnt 01 a more POlent.nd stable analOgua of
`GLP·2 (tOOl.
`
`Tha GlucagonJSacrallnlVu"acllva.lnla.Unal
`Paplida Family
`
`Chamoklnaa
`
`Peplkleo 01 this IanVII' e~t>er slan wilh TI"·AIa, HI.·
`Ala or His-Ser and are some1lmes p/lyslologlcall)o more
`imponam substtale.than whal OCIIJld be e.opec1ed lrom
`the substrate spec:;fcity 01 OPP IV.
`
`The plasma prOleasa responsible lor p'imary
`proleolytie cte.av.ge of growlh·hormone·rala..lng
`lactor (GRF) was sllown to be DPP IV, liberatl"ll T)'f(cid:173)
`Ala to give GRF (3.44).NH~. Thi. degrMalion Is
`responsirlle lor the ina.c1ivation 01 GAF both itlvirro and
`In 1li0'(l194,9SJ, and could be """reomo by inhitHtlon
`with dlprOl'" A, or b'f substilUlion 01 the Ilrst or second
`amino acod with a D·amin<> acid. A detailed kinelic
`aMlysis 01 DPP IV pr01&o1y$i. of GAF and analOgue.
`shOwed .ome Inwes'ing resu~. concerning the
`the Pt po.~ion, Values 01
`sub.trate .pa;;iticity at
`I<aoo'Km 10' dinerent P,-Sllbsl~U1ed aMIOg""" varied as
`IoIows : Abu > Pro > Ala » ser > GI)<. var » Leu,