SOME STUDIES ON ASPIRIN ESTERASEir.amu.ac.in/3329/1/DS 682.pdfsiCf y@t« l^
Transcript of SOME STUDIES ON ASPIRIN ESTERASEir.amu.ac.in/3329/1/DS 682.pdfsiCf y@t« l^
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SOME STUDIES ON ASPIRIN ESTERASE
DISSERTATION SUBMITTED IN PARTIAL FULFILMENT FOR THE DEGREE OF
MASTER OF PHILOSOPHY IN
BIOCHEMISTRY
BY
BIOCHEMISTRY DIVISION DEPARTMENT OF CHEMISTRY ALIGHRH MUSLIM UNIVERSITY
ALIOARH
1984
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CIIECKED'2002
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1
4* Aspiyjn aad I t s y r ia ia • ^Qet^rlaaH^ylie a&Jbd kiomm. popuiari./
as a j^ i r in i ^ ^ ) i s prolmbly tlsti ao«t wM#l^ \mmi dnj^ ifi t t ^ v#cHr-M«
eitliar aingly or M cocibiaatl^m witli 0tli@r drugs i i ^ t a ^ict Mupta*
1977)* I t 1^ t t e aoet ®ctei^iv@^ ^pli>^©d a^ali^alet i»tJ.|;^rr9tie
and ^Qti^inflaQ^atjary agia^t (Mato jg^ ^m, 11^2)* Al-y^^ugli a$lv< *9e
r u c t i o n s to iZa vme &re mn^wt timy ax^ nmv&rtiml&m ^^mm b@Gmtm&
Q£ the issdversal use of tiiis *^m&efmm*B m^lGinm^* M' ^B iy gj^ D4»»
1' 72ai« As a th^^e|ieutic a^#xit asiiiria presimts acN^tliing Q£ a
paradox. Jrhet lay!i@ii r e l i e s v^tm i t as tl t@ < ia@oa toiisidb«ilil «aalgfl^
siCf y@t« l^<^u^ tii@ dryg i s in@Kp«r)siv@ m& so Jtroe^ aimiial)l@9
lie Ql't^^i wad&CBt&B i t s aimlgesia efiica<^ it iOodlAiQry and Fiagl^
1: 73)* ^iiiice Um iiitSH^ductlcri of aspir in la tbm fiiddie ox' l^lti
eeitisryi, tiiere haim toe€» as aeyay reports oJalialtig surti efieatir©
anaXg^aia for i t itimi Go^parm witli aoditM saXieyiat@t as '%;lK.'r@
liav© bmm ^ios€ iliselaiGiinig si^p^rior anai^sdLa i(*ros@ sxid
yremiow^gp 'i':J^i aiit&t iij^$ ij^ua&m, l!;^!; i ijcon ^ lll«t 1^63|
Seed, 19ld).
i»'illo» faar^ Caali^ allmic "^^ ie^ti^yretic prc^erty c ' ^licti
was ^ m m to l^e axiGi^its* c^itaina a Mtt@r giyci^id@ call«El
niXioift. f i r s t diaooverea lir k^^QvoL m tS27* ^ i l i c in l i ^^mtes
glucose ajTKi s i l i c y l i c alc»3liol (aaleginin) cm l3(^drt»iy©is» For tim
f i r s t t i i ^ I'iz'ia (19JS), obtained sa l icy l ic &aM (orth^nliyclroxy
oaEisoic aoid) from aaliciri* ili@ syntJietlc imnufaotiire ai* tliis moid
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from gkw&ml waa aeooi^jjLislic^ in 1860 fc^ *ia3.ba Ciiia i.ciiit^«iMA,
ur&am*9 Mt rodue^ acetylsalioyULc aeM for therai.»eyi;ie imm M
tiie y^ir 1«i«>: « ^ l i o y i i c aeA4 i s v©ry irri"teatlag mm i t cati utiiy
be us<id ior siw'fac® api>ilcat4c«i« Va^'iaus derivatives Q£ UOM MiU,
iiave» timreforeg 'bowa oyziUmml^s^ ior &jstm^iM vmem i^^i^irm i s ai;
es ter Q£ sa i i cy l ic aoid In itiiich th@ carlK}ij(; X ^royp &r sol ic^i ic
acid i s i^tain@d arid lutbstittiticct i s male in tkm k^f^roxyl &s^iJ^ 'x^
aa a«:®tyl ^mtpm
jsi« 'Xli^rap^^tic ys#a of AsDirija • iiiere are laaay systemic a»d a
I'ev local %m<BB of tii© ^ilicylat@e. 3y^@ral are lias©^ on t ^ t r » l i *
t i tmal mid r ^ i r i c a l r e su l t s imtt^r ttmii mi a clsmt ijndi«nitai»dLiiig
Ql the iaeclja&iisiii oi' tl^rap^jitic li^u^i'it* Tim- pliari»€»ol&«^t aiKl
c l in ic ian ijmise tli@ efficacy aiid safety oi aspir in aa a» a»«ui^^ic
aiid antixiieii^iatic ag@nt| y@t tii@y fiiKl i t neeeasary to t ^ nei &Mm»
tant ly ol' i t s rul© ma a ^ iia^m caua@ of l@thal drug poisomns in
^oiCi^ milMr&i «AM se£*i£H4s " ^ ^ c i t y in imiylts i£ imM witt^ut
airport i.:i&(lical supsrvisiaa*
4aiiirin i s itxmUml^e xor t t o ncnspc^ifie x^lief oi c^rtaiii
types of pa i s , lik@ tm&^€ii@g aM ^liBi oss-ociatod witlbi artTiritiSt
iieus^l^ia a i^ £^lgia« Gla@aiieii ^ |^« i1S^2) Mv@ coi^aroa Um
eXioGt, Q£ tkm ic^px^ljosiate^aj^irin cos&iixitfitiim 4l-iicr@i£iiQ/ with
aspir in aloKie for re l ie f of EMMCIQ ctmtractioQ ttea^elie i^yC&i)*
Faticiit aM physician glottal ©v^D^tlox^ni supported the ccmclusitJti
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r&Hmrija^ the Bywgit&m Q&^I&K &£ mmelM &mtr&atimi b^mLaetam*
» cidcU»iry m^ Flngl (197^) i ev© r&pQrt^ that^ in the &as©
of a 3.i,rt8 r^ai£mt4e f€f\^r» the saliAylat^ii ®,i^€^a tli^ e^ti^tlv@
inflaiiiiat0i^ |ir^s@ss#s oi' the dJU^ase ibut do joot ejCf @ct the 'pro^
g^^t&imi o£ t ^ dis^^se cr the l a ^ r pitosi® o£ g/£wmii^m&tQ^m
!l;M£2mma.tJj(m Q^ scar toemitXimm D^^plte t l ^ <^w^#li^^nat oi mevm^
n@ver ais^s^tSt salleyJLates ar g s t i l l the &ea!nm%t€Hms> of t^ixni^ mm.
the drm^ ^£ f i r s t choice for the t rea ta i» t of xl3«iw&told a r th r i t i c
(B^aiiS g^ §^^9 1^^2)« In mMltSjm. to th@ mml&miA that alloiiB
stors @££€^tive thc^rapeutlc z e r o i s e s , tkmem i s iaproira@iat i^ appe
t i t e ami a feeling of well«4>eia^ ^alioflatins @1«& pedvom tkm
infl&tfiiaticm ojt ti^ J^int and siirrovsidi»g t ias i^s*
^^P« Sit S^* Ct97§)» havo preaaated avi4@&e@ that hi^yiitha*
s i s of irn'ostas^Landli^ 1PG£2* '^^^p*^ @s ^ ^ ^ ' orn i^Atelets ia a l e -
vatad ill vitasiizi S-4a£ieimit rmta* If th@ 0^iit3m&lB of petrntAgM^J^"
dina pla^s a si^ilfica^it ro la ia th@ d^v^lop^ent oX jpatholo^ In
ai^eciiic %lBsvm& of vita@Jji i)-<4«£iciaiat a^iaalsy th^t ebrujip Mii«ii
iuh ih i t tQ hltmyni^m^ia a i j ^ t aiieliox^te or ad£are^:^1^ t^ mml»
f astatioei o£ a vitamiii*£ 4ef ic iae(^ dep^KiJjig OB the oat^ans oi jmr-
t i cu l a r t isaue (HeXs^i ^ ^ J^ , t l^) i« i^|»iria ia a wall ia»jn«a
it^Oiibltor of FG s^ thaa i a ( V ^ , 1971} Vaoa, 1974| Flower« t^74i»
fha aaaooiatiim h e ^ ^ m afl^irin liica 4rvim ^ i ^ prostaglaaellas
hacasie cl^ti ' in 1971* hut @iricl€»ca had aiNsuaulatad iii the l a te
%IMIQB t a au^^est tliat aapir ia mad r#lmt@d lyati-'istfla^^tcary aji<mtB
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prostaglandin (Coiliv^'t t^Tt&i^)* Aspirin and related ant i*
isilMmmtoiy drugs Ija^rt^r© with i i l ^ m t i a n Qi iirostaglaiidins
from uplem% ieert^iim <^s^§ tWIi ami p la te i^ t s i^i i t l i ^od
t.lXlia» 1 ?1i mmk |ir@¥'aat@d t l ^ s^tli&aia of p^^taiA&DdiJis i^tom
arachidaciic acid iii t i s m ^ ^im&g&a&tm i^mrn^ 197t)»
A3|»irin i s vmM «ri4@i a s aa sutiei^MP^ijadat* ^ « veiiae of
aiitico&gulatifm «ritli dic»^sr^i miA ifsri^aia to avoid tlatwiboefflfaolie
a-offi^lieaticms in pat iants w i ^ t@(Ml i£^ to f&ra d o t s i 'iclsjuiirig
those with prostlietio hmrt vmXvas Ims 1^01 *^i i fi^ti^uULsfe^ iU^^^
msd iimmmt 1i?^i* M t i p l a t e l e t drugs mnoik s s a ^ i r l ^ and di^yridn-
mole have also b@an t Bod ©sct^isii^ly as alt@rsuatives to icsriaia
bocaua© tliey are safcrt Jjiaa^iiisiv©t @a^ to admiMatex* ami tim'g
require 110 la^oratoiy t e s t s Xar regulation (woinsteiii j | | ^ » 1'jS2^,
71^ InMliitof^ ef£@et o i low das@s o£ a ^ i r i a on p la to la t ai^^raga-
t ion tim led to s^vorml po t^ i t i a l t l israp^itic uses of thXs an%*
.voocitmry aiMi ^'ittgl C1975) Jjav© s-«it©d tli@ pc^opii^laotie adiaifiistra*
tior* of" asp i r i s ®g»rts a proteetive «Af©©t <^uivai«st t» tiiat oi*
oral astiooagulaiit a&^ats a ^ i a s t yf&^o/oB t liroia^sis and pulao^ary
eiBlmlism in patiefitc* u»dergoiiig c^rtalii ortlwapaodie op^rati^ms.
iii© S-Sii'j res©arcii grm^ (1'982) &«ive sli«««i ttiat dosage oi" aspir ia
used i s pt*o£^bly not dlffc^r^tt froe otliar <»ral ^itiocm^pilaists iii
@f lect ing siortality and i9orl»idit^ af t ^ a larocardial iataroticm*
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vjf
or eXisalimtea t i ie ti^^p@rcala@ffiia aa^^m la t ^ w i t i i ee r ta lu types of
ameer (^a^yfaertii ^ j ^ . , 1975) IM? Mitfe t J ^ adiiiJaldtrmtion of 1c<-
^Lmmmtruted Umt la rge doaea &£ aspdrin c^ys@ fc^irp^sgCle^dia i n
nm'm&l r a t s aCt@r % i&a%m^iim j ^ ||^«» t ^ O j ^Etd pO' g^uol&istrsitiQii
iU^ io ^ . j ^ t t ^ D * A s p i r i n ^[3ii p r o s t a ^ o l i n ( f ^ i ^ *^ ^ " ^ ^ ^ ^
aiimm tQ l ^ Q'lioctimi l i i carrac^tin^ aoa@ of tJam e t i o l o g i c a l
seqiielae o f diato^tea laeilitysCWaitzman j j | g ^ « tsi?7|V/aitzman, t979^«
Cm i^t©raa@s - iiStejrmses are "vmy vMtLLy ( U s t r i b u t s t i n anisial
t i ^aus i i . A t o t a l o f 33 iaulti|i>l&» fo r^s of solulxl^ ^t^raa@@ lMiv@
bei^ii r<as4diin^ ai'Mi tli@ occuxraiiiae o£ t t icn^ l o d i v i d t e ^ i:«»n»i i n itae
d iX fe rc i i t t i ss i j ^s Mt@rrelat@4 U ^ l i i ^ i mm liast@i*s» 1967>« Using
@p€sclfic substratiNi ^ M I l£ihiM.tox^ tb^a« i M t ^ s r o n ^ n ^ iiave l^ecn
€dMiract«iriai3cl l a t a £o%w msdn clasa&s| ear te^ los teras^Ay aryl@at«r*
au^@» dioXiii^ntermsas and a&&tyleat@f^ke@s* M l t ^ a c class^^ ar@
a lso l^t©£^og€iaiHiS|i t ^ i o l i M a earboi^leBtentstm ex ia t i n 1^
S0peimte I'orsts %gii3j» tli@ aryl4^t#ras@s» ofa^iliJD^it^msas send a i ^ r t y l *
@atei:*a@c s ar@ isioioi t o ooci£r i n % 10 aiMl ^ f^- 'os. r@@|jectlv@ly*
k im@b@r o i a&i<i:>ot Kla vMs l i i i i l^ i ib i t i m r i m ^ @@t@rms«a w l t l i
viaryii i i i d©4..re#s M Si>QCiXicit|', imve l>e®a used t » eliar®ct©ri»t»
©atex^ses* A H esteim3@@ liiaicii are rga^UJly in l i i l ^ i ted l i^ 4 i ia#pr^py i
f l o u r o iJiaspiMito i i i r l * ) mid paraoKOB laay w t i l | K ^ V © t© aiO-UUiit aij
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#s3@ftt4aiiy siiailar afcieijaiilsja o£ ri?aeticMi# Itoysveri, ©"iter t/^a
Q£ ^iitm-nsoa can al»o t)@ «listii%g^ii3h^« iiMrMgo i l ^ S ^ l b j lias
deacrilaed an mii^i» »r gro«i» of ©aayi^^i deai^iated as A-esturaaes
eaterastaa CtitiiAya© tkm lifiiroX^BlB of e a r ^ t ^ l esters aami as p -
liitroj^imiyl aeetat« mux piwsk:^X eee-tet^* Uiillkg 'Ui aiJL(^t^'aiNW«
s e n o ^ ©steras^s e m i b i t l i t t l e ac t iv i ty towaixis triimtyrifi and
are t»at Inhibited i^ i)Fr CKT paraosc^u infaet* tii^se ^ru^yiiea l^^-'c^
iyz© botfa Di'r aiMi garaoKOR rapidly iAldrliige, 19&3atbs Gouticjr,
I'^^^l l<li>imt@rt 19^) 9 alt^&u^i tiiey are 41sti»et xrc^i l^ie alalliyl-
f ^oro^iosphatase iiftiieli i^tdrolyz^ DFF but not ^a^&mssm or iiltr«>*
ph^tyl acetate (^toim^ir aiid Dliaa, 1^^)* M <»ii1 rmflit to t i ^ elioll-
nest@£«aaea and allentarmscsSg, A«>e t©xnBt3 i aiNS; inlilMt@<l i^ obelatlri^
a^aats ariu actlvmt^^ 1^ cal&lisa -umB i.'^imt@r» 1*i $. i^ertoii i^ni
Kal«»tf» 19^9)* ^mB0 findliiga s u r e s t tliat «@3t@]r ui@s ary oi^taLio-
erusyi^s i£^ ccnscr^pu^atl;^ t ^ atct^.nisfa Q£ t^ imtalytie actloei rni^
aiXi'er ii>imjat&r j ^ al>« 1: f>7)« l »r€V@r» ' c@ iKisle ipr^^ l < ' alalia^*
p^ valyea would ^ ^ to li@ inv^^lvod in tli0 l'i^cU'*olytlc r^aetltma
cataly^noL bff a l l i@ alxiim €^t@ras<^ iMmri^mm j ^ jyl^, 1' ^7|
BsrgiiiaiMi mitl tiimm^ 193B| ^^yQ.t©r j]^ j ^ » 19S7i* ^ ^ozyae or f i
nal ly imoMEi as E 600 estar%)s# ^^s £mm^ to b@ id&atX&al ^Xtki A*
©ateraiie \ 'ieii iJOiiiMtion stiMles y^lag o i s t l ^ l ^ma^X li^^pliate ^eru
k third type aJT eate^msi^ ^OiSjoh i^^roXyssm s i i ^ l e carci&ji^l
es ters Imve also te^i deaerl^gd (Bergioami g^ a l .« 19!^7| i^rg^aim
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1 i
a«a jiitotm, ij^jm tkmQe eatajmses a#al.gaai^ im C-ts^tertiaes, u^
not iiydroiyze *& r» aor ar^ tis©y lakilaited i^ Ajx 'i*. i » ©oati^ist -IA/
&B%mrmBmQ cma also lie elasaif iea i^ iliuJj'' aMl i ty to h^d£^»
i t i s BOt d ^ i r 'i/ taiej:* tlio a'l»illty tQ aat a t ai-i all«<^t^r ii^tctr*
faee jr@-4Uire3 a dUfer t^ t ca ta ly t ic ss^ianiaii* jriix^£r@ati€i iipane*
cboXasterol @st@raa@t a ^ lipoprotein li|2aae« th£@@ Q£ th& ^i^ysj^
^Mch ae t on ^lariagXy soluiile est^c^» x* i@iy^e the clK^liJ^sterasc^
aAu a l ies teraaes M t l ^ i r misc@ptll;»ility to InMoitloii i y |%ii%am>ti
(%ejni M JeL*» l ^ ^ ^ t ^ ) * ^ ^ ^4l)str^te ^ieci£iciti@a oX the
©steraae ia r a t j^n&r^iB and Irain kmv& h&^ atudimi kjxth Qr|. aa&*
j .aaplioi*m4g cocipmiiada aa seXeetive ©steir-aae imtlbltmm it^m^ Is^:^^,
iu^Og i t i s bcoomlxig clear tlmt ors3Si^pl^a|iM»'Qus €af%i<3yrtti& are
g,mmrmX inhibi tors for mm^&m poss^iaii^g WJtb&xyll& €3taraa« a c t i
v i ty , CtiyiaotxTpBli * trypaisif cti&lii^atM^ae ( t rue MMX pamMio)^
l i v e r estejmse did railk Upai^t acetyieateraae ar»d li-««at@rase as-@
iii^iibitud oy a variety oi" cofipcaigida l ike iJ ' 't tetxme1^^iylpyr&^»a-
pimta, tmjp^ppopyMlthiJm&s^frG^taoa^aAt^^ diph^oyl^^ajphfiiioc^iuUiri-
(late» 4jtc« a@verml %K>i ars Imva ^»»uii ^ m t varloubi fi^is&linmB%Brm3&h
af ter iiil'^Mtlosi ^^tli Dk"P cc^tainin^S^p 4.Q contain fiuiMijplwircHas
![Page 17: SOME STUDIES ON ASPIRIN ESTERASEir.amu.ac.in/3329/1/DS 682.pdfsiCf y@t« l^](https://reader033.fdocuments.in/reader033/viewer/2022041708/5e46a5e49e90cb4c882bc702/html5/thumbnails/17.jpg)
r-l
that r^ict ions %rltli m'gaiifi plttisjpiKSdroiii cc»|gmisids of sijc «it^rftaMi
jToilcm f i r s t o x ^ r kismtijom imd ar@ lilaeil®cii^Uir ia aature*
P« Aaaigia Eetgraae • 13isr# l e HJQ Gl«@ar pr ior i ty to t i ^ oba«r«6tica:i
&f an J g vl¥Q |}jf4rolysis of a ^ i r l a * 4lt^@i;^ I t MAS i»ot«l lif Lester
£ t j ^ * (1946) tiiat plai^as couM l^dr^l^se the drug, t ^ y ^ ¥ e mo data
aod tli@y did not i^^oailato If the |>rocesa ^mm masym&tU^ 'iim i^K>rt
oX V£»d#lli and S c a l t r i t l (1943) re&om^^s&A t t o ppmrnai^ of 6£i
acetyXaali&yXio ac^id ^ t e r ^ s e i a ssfVsraX t i s suss otii^* t^aam. biood»
^ ^ ^ ^ M M^ (1946) a lso ^Ki^Kistrated l^drolysis &i s sp i r ia lay
several t issues* ^m ma^pm t t o t i^talyi»i« the b^^t%^»iM Q£ aostya.-
salicyUUs aoM to sa l leyHc miid mui £r&a aes ta t s i s rmi<&tved to as
aspiriQ eater&ss iFl&are 1)* Mpirixi es t s r sss i s presast lia |ilsswi»
Xlffm" mid kidney jViBPidslli ai»i j i e s l t r i t i t ^ 3 ) « ^ttis preseno^ o£
t M s €ii @@ has 3l4M> Iteet r@p< rted in t t o gaati^Le ayKK a (iitaits*
cross jg^ 1 ^ , 1^77)t md BtmlX iMt&BtSmm C€iiZTy» 1977)« 3 ^ ra te oX
ooixv^rsioo of aspir in to ssMoylste J g irli^ @ay te Isportflot since
aspir in mm di££^e*@nt p r o p e r t i ^ to a s l i ^ l s t e (lisinsf^E^ j ^ jll^t
i^t^paes capaols o£ ^^drolys^ng s ^ i r i n are pras«nt in p^&mm
mid. variQtis otlter t i ssues and ma^ be iaport^Eit In dst^mining tim
plianmoological c eiid tojcicologieal propsrt iss o£ tiiis Onig,
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%#rg^#f ^ s^m^^mMmW
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COOH ^2°
.,OCOCHo\
Cv. Y
CH^OOOH COOH
-OH
Acetyl salicylic
acid
Acetyl Salicylic acid esterase
Salicylic acid
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J. J
especMUly 1 ^ dlam^ae s ta tes {ma^u^ jj^ j | | * » 'i)ff23^h$ Vlaeent
azud l,&ireu« 11Nt#i ri^rgapi said i : r u i t t » 196^} ieaadMisAr j | i y ^ t 1^77$
n&inBim^^ 19?7| i^aiasfom^ 1^73)* I t lias 1mm. mitimtte^ ^
tt<»flmi<i @mi ai«^@JLffiaEi (1963) t h a t a i ^ i r i a mKtt^rwi/m activity in Um
hiMM^ Mood volume aetK^nsitji f o r ab<»xt 20;^ d f t o t a l p a r a l y s i s ai: a
aas@ oi' a s p i r i n w i t h tim r©iml i i i£^ tak ing p;ia6Q M tim IXvmrm
fwme^^s^ emd i a e l » r (1975)9 ^ ^ w e&i^t^alatcd t l id ae t i t r i t y
o f a s p i r l a eatdmse i n plasiaa v i t b t l i s hf tM t i n t aad fmt# of « I i *
a inat icm ai «al ic3r lat« itod aiio«f«i t l i a t tlnim mis^aa ae t iv l ts f eouM
d«t®rcBJjte t ^ &oa»mpoiktmi&Qmi Gmx^mnixm o f as j^ i r ia t o «ft34;^late
M SiSBp ^^^»>9 «iuiek (1SM>6) and o*£r l« i i (196S> Imam elMMrvwl t l i a t
la^uyrJLous Gtt&ets oit ftoa^lJHyuui^jUUj aisid are iKMitJljf dua to tiMs
I n t a c t aolaeuXe i^th»r t^eKi t o i t a braaMowa j?r»iti»^t MK I poaai ls^
tha t ae^ f la f^s t i n a s p i r i n aataixiXiss a i ^ i » i i t r i l i u t a t o auoli
in^ to ' iaa . I t ^ast thsreiQx^^ asamtasLxy^ t o 4etei^i ia« afli^Uria
asteima@ a c t i v i t y i n a«ra o f t i ia mmm am oo^parad leLth tlioaa of
noi^^al Gontrola*
Asp i r i n ^ i to raaa dL££m^ J :*oa aeelgflehol i n ^ t ^ r a a a i n t t o t
i t i a 3 l i # i t l y I n l ^ l i i t o d t ^ ea«rina« i i ^ i b i t a d ^ laof loaropl iea*
l^iato (DFF), uaaffootod by ^^ani4@ ana caXoiuB ions t and fiaatrojrad
b^ b a i l i n g (Vincant ami iagrauy 1 9 4 9 ^ 1 9 ^ Qlassont 19^1 S M%iaraa»
19^2) Mid I t liaa b&mo, o lear3^ < l i f£u i*^ i t ia ted Srom t t o paauciooiio*
l ineaterasa o f sanis (Augustii isaont 19^$1SMi9f Baa t l4 i ^ 1d^>«
Xho r o l e of t h i a ani^^iie I n a a p i r i n aetatioliam lias l iMn r e o o ^ & i j ^
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mit i t ham o&t b@€» ado^pfttttlar tm&swc^ in animals auii m^
.>.,i f ^si2jca M aii>irla est&r^ase ac t iv i ty b&^»m«a mX&s md
f @aalds M aeimraX ^ 6 i ^ of ailsaals &M tssmn b@liig@ has been
reportad b^ s^vm^al Go«v€s*fcei Cl' aiMi ^ jlliai^ 1^72a)« Aetivit^r ti^is
found to l)@ isa^ Isv^r i a plasma i^ £^%u^ a^b^ects timet emk^
tliat ^ s t r i c uleei^tiaii ^@ to a ^ i r i n omim^^Ujsm i s aoi« prcfimieni
a^^oiig wimmi tbsm tmm^ ^ea aad lusoise t^^aeii arir aQr« a t ri«k
(Biilingtcait 1963$ mmsm sod Ctaapiaati, 1970)* AajK» i t mM ob&mrvmd
|}^ Collina and furxisr il97^) t ^ t tli«ar« M ^ ai^ iii er^ftsflcl i2iGi4aii«e
e t i^teiti^l m^mi&9 mktm^^pmrtmt imxwspiim^m^ ppelongad i s s ta t ion ,
6te» i& a gix»^ &£ pstw&MEsxt imwmi «^o %fed iisiiii^uai wu^a o£ ae@%i<»
saiid^flie mid. m^w sm e&*m&eki^n i^gimF £1 A^9 1^7i^»^;
Vinoent 313d iMg^em^^ l ^ ^ i Itoi^iii «ia ^^niitt» 19^ ) tittve mggwted
tliat tJbe 4m»rm^iJim in piaaiift asi)iria i^texmas n^ai^ they <A6«r«@^
i a i»c @% mad p a t i ^ i t s itfitli ai<^^o3Jle Xiv^r diMoam m^ be relatod
to tfyts iia^roessd pc^^i^fti'^ o£ t b ^ e two grcnii^ to gas t r ic ttl«tii«*
xSjm mid ^s^maaerimm* ^ ^ r^pc»rt^ di££&mxGi9» i a 9l««aft aipirii^
«std»yie im^mea t^ msmi md M pati^mts wXth alcoiK>Iic l iv^r
disdase ^mrm v&*ixffnmtl&k$i9d ^m& a iso pa t i ^ i t s with siimamtQid
a r t h r i t i s im^m l3efi» stysiiad* Xlia l a t t e r grrn^ oojq;iri«a aa a^k^raei*
alxl« pEt^partioii of tlia popiiiatii», r^quis^ hS^ <iiMM» G£ aspirin.
and. are immm to tmv® dafeeta in iiv^:* m&t^lmllm& i&sndail j j^ j j ^»
1d?1| HiitGii g^ ^ , 1974) wiiioh a>uid ra f lae t tha ^ p a a i t y of mis
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n
oruan to pi^oduce ©azyaes iimroivcd ia l^y«lroiysiiito asplr to ii- i -i%-
t^iatice a i^ iwth ( I ' ^ t ) toave oiimas *yiat a l a r ^ i ract ieu QI
tii© aspirisi ostermse i^ t lv i ty ©i t/tole blood cmiM be ifiK^tad ii'i mc
tiie ^i^e@ levels in &c)ftka'QQy%m and plaai^ mit^estiji^ Um% tiie
^ ij ' e ^)3id@ lii^ t^ c e l l ^ad plasaii ^ty "m aX£tmrmL%m 'i>l»@ i ^ l l y
l a r miz^wm ia probai^y a nilatlv^J^r msm^mciSUi @sli©rsia# ^@r@as
toe platiiia &&z^'vm ^^t lvi ty la di^ ta c^lM^t.@raa& UB -gs^wik^^mlf
Scoi^ of the ^SNsey t 'liork - iispirln i s tl»@ aost wictoly wsed
aaalg@3iCy C3a:itipyr@tie a^d anti^infla^mtor^ drug a^d l)Qt i usex'ul
Mui toxic el'f seta af the dnag mre attrilAsted to ti]^ l»tact saoleoiile.
Hy^ndlyaiis of aspir in i s ca t a J^z^ \y est@x ijii3s present iri imiuous
t i a s i ^ s . I^w@yer, 'Hie blood aspir in ^t^tras© i^s not beaa fully
diaract&rised* tium object, of tlte priis€^t i^ark i s to tikmro&t&tlssm
V^ principle @D>2ymiiu} resiHSsisll^ t&c aspir in taydrolysis in M.oo4«
i'^st Of t i ^ aspir in i^srdrcly:£i»g ac t iv i ty oX t ba tilo<^ r@aides la
r#d lilood ceHs but s^ far no nys to^c s t ^ y ia iiwailalile oe th©
cthai:'Ectai*latics Q£ tl^e r ^ M^KKI c e l l «m:^raes« 'lite present %iork
repres^trws ^ a t t e i ^ t to d iarac t t r i so tiM} rod lalocKi ceH a ^ l r l n
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wmm:^mL
![Page 24: SOME STUDIES ON ASPIRIN ESTERASEir.amu.ac.in/3329/1/DS 682.pdfsiCf y@t« l^](https://reader033.fdocuments.in/reader033/viewer/2022041708/5e46a5e49e90cb4c882bc702/html5/thumbnails/24.jpg)
± IJ
!m@ Gb@&<iimJLs vmeA i a tlie p;*@8€nt stucUUfl were ol»tai2a«il
SFma tkm mmpcmi e$vm. n i a l s s t the i r mt&mm O t ^ r mmke&lB nmt^
v&pQ o£ msHytlGBX igcs^» or t t e best 0p&^ 4Mmlliil>l# ooaasreiaU^.
«SBSaSSSSSSSSS3BBK
Bovine Q&nm oXUmM
Q st@JLQ© £9«lroehlisr*Me
Etl^leoe dieusin® t@tim a«@tate
f &rric ititrate
M&K\MPXG d i lorMe
a i l fu r ie i^sM
f r i s (liydrax^iaetiiyl)
B«ll*E«» India*
India*
B*&*li«^ India*
Surailiiai M* Qtffialiml>> lniflSa»
B*o*ll*, India*
Mg^a ai@iaicaX Qci* y*^*A*
u*«.»» A«
B*iJ*li«p India*
B«;>*ll*« India*
B*B.ii*» India*
^•iJ*M*9 India*
![Page 25: SOME STUDIES ON ASPIRIN ESTERASEir.amu.ac.in/3329/1/DS 682.pdfsiCf y@t« l^](https://reader033.fdocuments.in/reader033/viewer/2022041708/5e46a5e49e90cb4c882bc702/html5/thumbnails/25.jpg)
14
Xt i^a » 2s@%)iei2t2y cso Led eM t O i^ <liatt-lled watia' was aiM@d«
i%@ pr#cl|>itated aspirin i us £ilter@4 'y»r<<»i & B^^midr SvamU
t^ m&piriM ^r^paxmtiGB was p u r i f i ^ tj^ n^«ittc4 cgygt«lliiftt4<m
^mm # Agglrte i t ^ m a ^ - ^ ^ i r i a ^t^?iifl# ^ ^ v i ^ m bii»©<i @j3d otli©r tismi^s «as ^tesm$ii&d as dy@scril}6d lay C a a and ( ipti
(1977)* fMs i s a modiifieatioa of t l^ |« dOsdisr@ 4meeib9A h^
fria^«» C1:?§4).
aflpiriza solutic»k (300 sis soiialja^ aspirin dissi^ipsd la KK) ml 0*06
^ !l ris«tiCi biiff®p« |ili 7*49 vm& ^ • s s ^ ^rsj^arsd inBatdiataly liaftxre
1180), meid 2,0 ai of 0*06 j | tOris-ftOI lKiff«»» pH 7*4» ^ a aiimra
iiaa imiaiat0d f i»> ^ M^»it®s at ^
i aaklzig* 5«3 sO. <^ aoldie«fi^a^c(IKij)» 8#l$^iQB i«ts tlMH added to
aaoh tuOie aiid tlie isoiitaats liei^ agltatad irig)S»«ysl^» oaei^r'ifi^id
at KK> r.p«a, for 13 aimites* Ce3.otir of Hi^ supaflaataat iias raaci
iiiaediatal^ at 340 i ^ a^tiast a r^tg€i2t I Xaisk* mm fld^MMriKoioe as
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15
carreotod e^bsorbrngm A^CA^-A^) for eacb sasple «as « i«» iis«d imr
graiik* a m i sftlicylifi ai^^ ««» u^d for tlie prsqpuimtlMi «€ a
sti^f^tf^ airve» i^jplria ^it^rase ws^U^l^ Is «9 ps*«sa«& M I inaJUw
!llie ooilovs* roa^efi't luiedL in tlie aa^a^ vaa iMrasiemi iBf dlasoXi"^
jjfig 10 m smF&xpiQ €liljOri4e in I6a<»t70 s£k b&t i l iatl l lad Mat«r» Sli©
soOutim «fas coolad aQd6O£aIoi: i | | iMXlaii«l lO0a f•ilK^^)*SWi^ w&e
jatMyi si ban ftlH tib@ t&^iM £iltyra%a IKBMI dljM(M^vwl« ^ ^ voliaw was
w^s» \3$i^ ^ 0 &Jl ultlt 44atiJLlMl iia'ter* 1 ^ ri^^am^ cotildi INI atc ced
JjMlafi iait^^*
4lRR^4fi i i l i P l i l t mM^ jfe i l^g^ * C itdated ismm^ tOood
ol^taified f r i ^ tl)@ 3^oal llaSi4^ CdlJu^e <r i r lli liilooft dswatad Iqr
^ e a l t ^ -voXtjiita^^ was vms&m B^ blood eaJila s&A pk&mm wmm ai^a*
ratad ts^r ceatr i fu^t iaa aa^ tba red laood ^ULs W K ^ naaiiad MW^ml
tism& wltli nonaaX saliaa* ^ e r d ^ to sti3^ tim af£eet of trnpara-
tura Mid pH on tlie aetivi-ty of aspirin asti^'aa^ rad bleed eelX
hm&2^smt& (rad Mood ooll t dlat l l lad «at^*» t t4) wtm pe^^^ucwA^
A9Pcois»iate ^loimt of tlie li^^lysata i«a talcea la tbe assay olsmtrt^
to oarr^ out Van^m asq^ariaaeiits at imrimm ^ moA tooparatojraa* Fcr
tim Sx^^iiMtxiry 8l»ai i^ on thm rad lilood oelX ani pMit ai^^afofrlata
asaauota of oitrata* ptioi^^ta m' iodoi»»atat€i iiara JiaaliaSad ia tbm
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lb
with tb@ tmrlcHia coppeunda lot* ten ulsutes tieimm tibm commoiemmxt
hypotofOe 8oauti<i» C5 Hi fria-uca tstcTf r, |ii 7,4) WHC ussO*
a ^ t t ^ r Elveh^aift lioasgiiaiz^r with & xm>m& f i i t iair f«itXiiB l^Mtl«»
1 ^ % ¥ lifinaegeiiat^ iiffif*e pr^j^ara^ He^apgatsatga ei: vnri<»ift tiasuBs
tli£! sl^t^iEiii^tifm oit aap^ia it r!&&& ^el.lylti'*
study msiRijg ikfid aidMlt GUMB mE4s@ staieed £gmt Itx^^mi VcterliiAiy
H#9 ii7« i It3»stl1»j(l€,» lzmtKm^;p (y«i «-) i#eic^ »e4»a* 1 «s ts^ljaala tNara
maintained (m stancteM liiii{|iMt^& l4svtx' r&t/miiim £m&m Tkm yomm
and a iult aaimls ^®re divMM Into 4 sro^s» i^l2« 1^ eentroi
r mog j ^ ^ a^at wfrmjip» watSmmd t e Peet^ivit tli® imml 4ia%9 ^ «
iale& &X t r e a t s ^cm^s jr«e#lv@d 1 ^ ^iEg teo^ ifftl^i/^ar aaplrin
in «li«t for 3 w^^« Cars was tak^ te a@@ tii&t tii@ miee canttaagBl
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1 7 ± 5
a l l t ^ aspirin wiiieli warn nlvett atlaEod in a aaali aaomt of 4 i * t *
levels of aspirin dsteredi» w^r« seGtsuriid In tb& hoiKigttiiatoa &£
Hi^^^y gastric massfmm,^ Ikw&t m^ inteat^id*
method <%£ Bradl£<3rdi ( t9l^)t tynlG^ oocNisyBKsit IdAMi ^ S HOs* BCMRUM
s rum aUnsaln WEUI iH^di as st CKlardl £f^ tlMs jpipi pax liiQii o£ ^liba^—
t r l U i ^ t Wxm a«> C} in ^ ca ooe ^ $ <9^^1 al«s€ib&l« 1€0 a l of S ^
Cv/v) «srt ti ^^d8l i6rie ^ i a sas acldail t^ IMa s@l;Ht>i«i» Host of tii©
dl'e diafiolima l i |ilieff|ti6rie a eM and iSm mlmsst af t ^ llr« also
to & f ina l i^linid a:£ ee® 14t£%*
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H M y i ^
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u
act l i r l% of }xamm Ikl^od a^^i^tmm t o r^O^a b o ^ l a red blood ^ U s
and pOasBta* MaJjfsJLs «^ tfa© S^laaia f^^iltod in a s X i ^ t lii6r«&8«
i a tiie aotlYity (fable Xi*
Most oftjift noiic OB aspi r in ®at&p&»& kma IMMB os^^iad out
using blood* plaasa or ^s*ua aa the aem^e (sae t^saii^utord j ^ §^9
Vjm^m c:ostailo m^ G^em H'MZ) reported tiiat r ^ l»Iood o^lla
ai^iibit al^yif i^ai^t a ^ l r i a t^^clrolyzlng a^feivlt^* we liave alao
Qb&mr^&A timt aspi r ia aet^maa £iotiirit^ of the rod b24>od eeUa ia
Gcmsideralaly Mgbar '^laii altl^er plaaaa or s^i j^* ^ aboiA ia
faoXe I t a ^ i r l a ^ t ^ r a a e ac t iv i ty was al)oyt a*£old iililiar Ui red
blood ceHa as ootipared to ^lasiaa sm& ab«2t 4 t l^ea liliber Vami
wlKila b lo (^«
In tl3i@ r@d lilood emUs mii^ a s^al l Amotion of a^M^ln
63t@raa@ aiCtlvity ims £mm^ to b@ aasooiated t i l ^ tbm awdaraoe.
Aa e^ii^iPtt JDD^I l^bla l l * exteioaivel^ »aals^d gboat eal la ooeitaia
onJ^ about t jp ot tbe a ^ l r l a ^stax^sa a o t l r i l ^ tdiHa aver 7!^ o£
t i » act ivi ty could lie loealiaad l a tbe @^^reiia<*£ree l^i^Milysate*
As also 3imm In t t e tat^Lot tba i ^ t iv i ty Q£ aspir in ^utarase «aa
found to be M^Oer la beiaolysed red blood eel ls*
Ttim r@d blood (^11 hmaolyz&t& «diiblted two ei^pirla estereae
ac t iv i l ^ pmkB vAt i studied aa a ftxiotioa o£ pii (Figur© 2)* Hie
tXrat peak aisd ac t iv i ty was obeM*ved a t lii 5«^t tii^reas tbe aore
^tmaismxt and re la t ively sharp peali yas located a t 9H a*3« Xhese
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19
^WA - I
ijjia.tritoytioaa of MpirAa psteraae Actlyltf i a Blood - ereshXy obtaimNi eitr^ttad Caiman M o ^ mk» oemtrXSv^ed t o separate plasaa fro^a red blood cel ls* ajdtable aH^pot of {yass^ was dialyEQd againat 0.06 | i TriS'4iCl buffer, pB 7«4. Activity of th@ wl l@ blcH>dt plai^Ki, ^ s i i ^ r@d l»Iood ce l l s ar^l dialjrzed plasiia t#aB aa ta r^ i i i ^ as described in tim %^&tm
^urcB Aspirin ©st^rmse actiirity
i\mXt&/ml) Activity
ik'hOle blOCKi
Flassia
Had blood c e l l s
Dia ly^od plMBma^
Q.m
0,23
2.06
0.40
1CX3.Q0
49.00
403.00
78.00
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Zil
fable • IX
in t ac t r@d IsXoi^ «$@Xl3« ima^armm fiNse Ite^ilyasate arid ^ o s t cexis wer@ prepar^a as desar-ibi^ £iiid aapirin «sit^ras« act ivi ty ^nas d@tm%sdxtMm MmxJUm^ ac t iv i ty obtaiAdd la i a t ac t red bJLo€»d Q®11M was talc€si ms 100 to caaX^iXat^ tkm p^temt ac t iv i ty i» other sasspXea*
Source Aspirizi ©st@ras@ ^ Activity (units/ml)
Intact r ^ blood 3.71 1CX).CW cells
I'ie^raia© fre© 2,25 7f?«0C> hei^lysate
Oiost cells 0,49 1 3 . ^
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Figure- . Effect o£ m m tkm Aetivity of
tlwmiB. ii&^ B3.QOA CeXl Aspixla
Aspirin ^is prepared In )3ui£@rs ol iiidieated ptl* Using tliase s^alyti<ms of aspirin» activity oi' aspirin esterase V^B determined as described ixi the text* Tbe buffers used for various p« ranges were sodiiiui acetate a M 'iris-HCl tmffer, llm laoiarity o£ all buffer solutions \iias 0«a« Activity at pH 3*^ %ii talcum as 100^ to calculate tlie activity at ot^r i i values*
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:in »
rti.)
6 i i
20
. ^ j ™ 4 .
3 A 5 6 7 8 9
PH
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7.
s^icUes ixKlicate t i m t s ^ i r i a ij^Oroa^zIng a c t i v i t y a f the r ^
bioc»4 c # l i @ay i a i r ^ v e aore tbai i one en^@e« a i a i l a r lafer@iiee
cayQ l3e a ^ e £roa F i ^ i z ^ 3 M ^:iijc;b ftspirin iJ^fdz^lyzlng a c t i v i t y
«^s iii^^ujradl as a l^sieticm aX t@i^^»«umti;re* A re ia t iveXy ahaxp
a c t i v i t y p@ak i^s cl»3er9@(i a t 40^ nMc^ ^ ^ i i i i e d a t iiii^ber t « i i e r a *
t tB^« ISier© ««a8« iioii^v^r-t ^ i r i do i t a ab^mldar a t ^S^*
' H ^ e f f e c t o f l i o i ^ OI»^»OIM:»IS <m ajsptr in a«t@raji« i s deser ib -
@d i n S'aoie I I I . 'Skm i n h i b i t i o n of a s p i r i n ^ i l r o i y z i i i ^ a c t i v i t y i i i
reiE^ponae t o c i t r a t e wmA ^imm^m^ uiui iaaa @a£ika4 l a <mae of iiteoia
blood as ocM paJE ^ t o %i €» jri@A yiood o e i i s «i r@ usad as tta^ ^^ i reo
o£ th© @»s!y@e« i'ha 2^KI l i iood eeiX eo^a^ne ym» i x^ i ib i tad 1s^ io^loaca-
tamide*
^* jglfff^ 914ffff^lii, fiffiWfi ffi 1 ^ i waayrla iSi^arnt #ttyal^ i l y ^ ^ f i ^ ^ tf^ I f j ^ «» i n ortler t o 3t»«dy tha ^ f a e t o£ a^;»ir in £aa&*
i n g t young (12 w&^m} and a^ i i l t (3^ 'm&s^ml oaia aniiaal.i ii«r% aa l^ r t -
aO* fhe aniiaala i f^ra givah t ^ sagAs )^^# « e i ^ t a s p i r i n mlxwl i n
4 ia t« 4 t the €x^ of 3 liec^ssp tha anlBiels « » « aai»*ifi0«& «»d
a c t i v i t y o f aapir l ja ^ t a r a a a (tetarmiriadi l a ki4Bfl|^t i i v ^ 9 gas t r i c
Micosa msSi^ iatast iEia ( l a b i a IV )• SSu r \m& a s i g n i f i c s f i t d a c i ^ ^
o f aap i r i a c^toraaa ac t i v i t ^ r I n cmae o f IdUloey and ga^^r ic imicosa
o f the aap i r i a fad asiiaals* I M s vas oi»a®rved both i n ycnaig aaod
adu l t i3ic©« I n c<mtraat« the p a c i f i c a c t i v i t y of Hv^:* mA i n t e s -
t i n o miK/sm ^aaa aoc^s^mt h igbar i n a ^ i r i n fad aic@»
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'Hy^ asp i r ia esterase ac t iv i ty miM mmmur^ under standartl c(mditifi«i® a t various t 2p@x <-> tu res . Ao t i^ ty obtained a t 40* %ia3 ta^ffi as 1C^ to caleu*-i a t e th@ percent ae t iv i ty a t o t ^ r t@@|»@imtured*
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100
so
o <
o
60 [
40
20 F
20 40 60 80
Temperature (**C )
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9 /5
^\3im • I I I
•^^i^™ifcwfc ^FiWB^iWMfr
l«l»ole M e ^ i i M tilood oella % Aet iv i ty
100.00
aa.c^
N.0*
iuli*
100,00
62.00
43.00
40.00
Citrate 10
BmQ0&te 10
Iod<^c@tamMe 10
ll.l>« • n e t <leteinsM@^.
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g
! 3
g 8
I 38
#5*
o
I €4
tfV
«fi b* f -* • iMi f»
• • j »
o o o • t •§ • •
^ s n d o*
<J o* 44 4 t 44
3 S i <?
« » ^ o o o •••I «•! 44
9 ^ R o o o
©• o* d • i •§ • •
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df ^
s o 4«
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o 44 O m
4
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hj '0
a c t i v i t y JLn mms tismmB d£ a ! / ^ , im fc^ t f sheep asxl ImlXalo* t iMs
ej^>«r l j i i^ t Idas uodertia&api w i t h a view te f i n d out a Gm^mSmiX muk
r i c h i^Hiirce f o r tim iRfirificatJUm ai3d o o ^ ^ r a t i t ^ stud^ o f a s p i r i n
aaterase* As evident froia l^ie tafele* tli© m^^mm i uc t i v i t y wui fcyuod
t o h& h i^ber iA k i t ^ i ^ and Xiv&r o f vorimm mkXmaiM^ i d v u * of t l ia
ra&b i t @JCliiMted M g l ^ t n^ieoif i c a c t i v i t y auseogst t l ia t iaauga of
t i ie varioua an is^ ia l i a r s ^ t i ^ t odU
For tim ^aka o f conpariaon w i t l i t i ^ tuamsa maatyam mum
ai^perim^ita on the aati^ra of t l ia a h e ^ Mvi^r «B23f£io wore aiao pw^
iortm&m ^ ^vonm in Figure 4 a ^ i r i a ^ i t ^ m a a a c t i v i t y aatliifaitaci
a p@e^ a t ptt 6* t l i a @ff aet of t ^ ^ r i ^ t t i r a on t l ie anp i r i n aat^raaa
a c t i v i t y i s ^}^iya I n F i ^ a ^ % jkha syBas^ M x ^ i s ^ «@a ix^Gv^mted ixo'
^ aimxtaa a t var i i» is t^^iiejf^ttares* IMkiair tbesa ooia i i i t io i^ the
te^mKjf^tiM*a optiraet %iaa fmind t o be %f^m At €0^, tba «fia;y<Eae a c t i v i t y
was aiosaat n a g i i g i o i a *
%^Q a f f e c t o f vmrio«si mi i^^ inces oo t l ia a ^ i i r i n l^pdroiysiag
a c t i v i t y i s £»ii]mariaad i n tmMm Vl» y^-iaiNro^ptiMitliaiacJ. oo^ etISQriane
d iaaina t e t r a acatata produced no s i ^ n i f i c ^ d t @f f ac t t but aooe
atiim^dation o f the a c t i v i t y imm (^sarvad I n ffpmmBnom o f cyata lna*
i l C i *
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dS»j r.ii
5 I
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o •Ha at
4* ,1
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m
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Hd
6 1 1
8 T
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Aspirin mm prepared in tmf Jt^v of indieat^ 1 ^ Uain^ tbmm mlv^mm of aspirliti aot iv i^ ot a ^ iria as^rase was det^naiaed as daseribsd in the ta:»ct» Th9 buffers used for various pii revises wti^ sodiim a^sitata and frisoHCl &uffi « th& m&larlty of a l l buffar solutions was 0»2« Aotivi'^ at pH 6«0 wm takan as 100^ to oaleu* late tli@ activity at otiier pH valiiea*
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Fl©ir©-5» Biimt ^£ M ^ „ M^t^%¥
'Dm mspiria. est^xmse iMStlirity MNMmr xmdmt st^n^aM '&(mditi<Mis a t various tmsiem^tmBwam Activity abtalii* 0d a t ^ ^ was tal£«» as tOQpi to oalcu* Mte IM» pmremit activity a t ottor
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100
:^ QOt
o o
E 60(
£ X
o 401-
201-
20 40 60
Temperature (®C)
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Taiae - VI
P f f ^ ? ^^ yarlotts Si^stances m the Aaplrto m&^^Xj^R Actl.vi.t¥, ^ m i a s s a ^ ^ respects to j:^ag©ati$ ^irea beXow and sa^rsie acwrit^r laas
ilMitioa m % Activity
Hone ^ 1(K)»00
j^-ffiercapto0thaaol 10 8 4 * ^
Cysteiae^Cl 10 11^.00
Etl^laio diaioine 10 ^ . 0 0 t e t r a acetate
30
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1
u@ai < mg ksi&m JMtQ mt :^>* 1S82)« Bespit© -^a istsogsmii ^ i t
aspirin % a^ir i i^ aateras© iii iti^ m^ o^i^r ap^ies^
m@ .JE ^^^0 "ImmX of -aspirin. % d@% r@ @& % s'S^e^al
f^6toi?a«. i&eXii9li2^ a^&s^tifin mt@ s^ ' tli© rats c^'ms^^i^Mi& l^tr^*'
Ei#i©3£iasi« 19^<| Bo ia23 . ^ ^.^ 1972>« .ioi^ @iitl2&f &eM^@ Wm^
benoSl'&i&X effects of aspirin are diio to t l ^ lataiet ia&l !m33# mlf
effects ar@ ise^ateS ^ a ^msi^ri pitla^a^ lisv^itrJ^ 1© 4 n ; ^ ^ f^^ t
on prostafiandin Ms^ps^^sis ao^or x X@as@ (V@zi@t if?6}* QiaS^
(1966) azi€ O^Bri st (ISiS) haw ta1»M i@<l l^at ^la ^ i ^ ^ l ^ of
a&et^ismli@yli6 aei^ id 4\i@ to tim inlac^. ssii^ei^' zmtl^r '^m. i t s
lE^'la^^tes*. SH^ af^irin estsi^i^e a&tii^% in ,@@r -of -sm^ ci^r^aio
%is~ i isaf fu:rtl!S3 '' aj^tm&t® th^ cc^^in^ ^fse ts , dt M#i. int@li@ ^3^
@f @^ g^elat^ def i ^ In aspirin {setabo liam C^ t ta mM (^tm^ 1i?7)
4 t t^g nasse t;^^* i t i3 |}^^^i^ie Wskt ^ e 1@ ^^s^© a^tivi*^ eeuld
^nd w^"^^ ^^^ i i ^ iMt th@ en^r^^ aotivi% ^^SSB
circle l^ydisg to vsarieus izi jS'i s in tl}@ ^^ircsii^ laaer^
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32
buftloa aM izature «f aspirin ©steams©- i^ Ixti Ei layUse aM tismtg^
b&B ^m earrlei out itsiog pji sma or $@F«:BB a@ # ^ emsre© (aais^if^^^
that wliole biooa cejr ao&taSa relatively M^li©r i^msit of aspilj?!!
t9?1)« M sh.9im i£i l^iO^ I , m^B% ef "^e aotivl% of tt^ @liol@ ^oo^
aps^ars to re@i4f <:. ij& rs^ lalood 6#3Ja» H^lBssm. <sai>itaii»^ €Qisf t ^
of thi^ 'activity ^licl^ isms' activated sOl^t iy as a s^esult of dlal^*
a is^ Jdialysia of tlm plasma ^m tmrl^ out 4». vXm &£ oiir o^^^va*
ticm 1 2at sois© or asTii aoi^ss inhibit i^ mz^^^ @otivi%«
$^i@ II9 s ( is& i^ t to he ^sooiat@«l'. 1^^. ttta^ m^^smm^ ^NIs d a s r
iy .sm^m^ "^mt a ^ i r i a l^i^rol^la i s m^t- ^m to t2i@' ^SN^^UE^^^IS^IS©
jM<^ ia .aa©ooi&t@d with ^ te. r<^ l»3U}6<i €ci|U.- ii^ii^^^^*: A m^iX
tMa ,aatl%'ity c e i ^ -mt hQ -^i^^2M^f''^M
£lea.^$ 3S|>iri^ C^ofstee'# 19^1)« f s i ^ ^ -^l i i i l^s eti^. : ^^ : ^ r t ^@
^<l^i^'3^^n;g aetl '^it^^ sga 'aela t l ^ t «t£ Si E iffi*. I t |ie€ . ^L&^ I s^d 7®por*
th& disappsax^mce of aspir^. firosi t ^ . 'S^L& 1^^^ ^ n?^^ il«@@iai #
196iS>«. Our ppeiiakiai^ s t ^ i s s l&iliGate ^m.% ^20 i^^S'saa^ of tfe©.
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33
red WLQQ^ mXS^ mi^ s^eatx'iot
tiiiQler the asaa^ e@Mltt«:i@
mam ^c^n <m3 ^ ^ ^ * i ^ :&£ ^ e r ^ ^@od ^^J^» . a^ ias i^ 1 ^ - 1 ^
%9rl ^ m?Mi^t .teem Wtm- isM-$MmBl;p. #^g$Jy^et a s ^ ' ^ t ^ - fsa^ ^
c i t r a t e 582 l ^ss^Js©% si:g»i^i<sss5tif *^^t3?i«t a '. i?$a-
«f
a% ^^^*
XU,
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34
aspirin ©s'^ras© ii^ tsm t^mmm #f imriotis MJi aX©- m^ Jb i as
s^'^leets (Morgisi m^ fruitty. tSi^)* M@ Mir© i^^r^ier imrg'Sti^titil
til© actltrity in t^ various tissues of mle^ and jr^Mts (fal)!© V)*
1943| Fi?a%©lla astt ,^»®ila» t tS3 | ^ i t ec ress gg ^•.# 1$77) lisivt
gas^ lc i!ai€ @€t* ^ t@ activity o£ ^#piria ^t^^t^^® ^as naii^ M#©r $m
the l i irer of ^eej^ mxd iaai^ts^o^ fhs^ mm^ XS.'vm' mi^:0im QXJ$6 %WP®^^
to 1}@ a ^ l o X r@(|isiriJ3g ^)^m@ (^l}I@ ¥t)* 13i@ aii^ t E^sx l a:*®
wms £>l>;s@r@d with -^a she@p l i v ^ •m:^wm B^.g@sting t@ i&n^l^spiisit
of a siiigl© ©tj M©* aJhese -^^©rijgsats w$r© \p^rfoai^^d Sa ^ ^ ©f tN
very lii#3 ac t iv i ty © ^@ mii^m i a th@ ig^©i l iv^? ^bXi^ ap^tara
t o y@< as a #£»od BOUS C© £&r th© ftj^lfieatic^ of ih^ ms^mB.»
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35
QR^ms la waSxkky r e s t r i c t s t& ^fHam&m^ Bm^nor^ &mm r©port$
an4 data preua^t In tlia dissertation isedieat® t ^ t t$i@ r^
blood eells laay p>Xay a most ieportazit r&le 4> t ^ ]^dro2.ysis
o£ asj^ln» Attespta wiXl^ therefore* 1»@ ®de to JL^late m&
pwpiS^ to hoss>g€Q©i'ly t^& hmBsx @^ytta^;^te o ^ i r i a esterase*
D@taili»d studies mi ^m i^ysical^ yi^ustie and Mologi^^ |{ro<»
pertias M i l be iz dd* ^©oific antibodies ^ i l l b@ raised ag^iiist
-^e purif 1 ^ a ^ i r i n est©ras© to datermin® tb^ tmrnlass^ of th^
red bloM c@ll» senm md eos^toe from -mriotm o^^^ tissises*
In vi@%r of the lino^si di£f€:i»aaces M a^ i r i s i est@ras# a c ^ v i ^
bet^eea nalss and f emtles of several axiiiml species m& tsmm
b0iag3» i t liTill alao be of interest to study the ^rosaareaotlvlty
of the ensyiid isolated ^'o® mm and ois sn Q3i»s antibodies raisi^d
agoifiat en2^me deriir<§d from w^ ^msrc®^
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BiBummAmn
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38
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3?
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