American Journal of Drug Discovery and Development, 2013

ISSN2150!2"# $ D%I& 10'3(23$a)dd'2013'

* 2013 Academic Journals Inc'
Antimicro+ial Activity of ,-ytoc-emicals .#tracted from
Acant-us ilicifolius /eaves Against Stap-ylococcus aureus& An In
silico Approac-
0anes- Se1aran, J' Jannet 2ennila and Annie 3ercy Aal 4as1ar
Department of 4ioinformatics, Sc-ool of 4iotec-nology and 5ealt- Sciences, 6arunya 7niversity, 8!111!,
9oim+atore, India
9orresponding Aut-or& Dr' J' Jannet 2ennila, Department of4ioinformatics, 6arunya 7niversity, 9oim+atore8!111!,
:amilNadu, India
A4S:;A9:
Acant-us ilicifolius is lesser1no<n medicinal plant is used as medicine in traditional systems'
Stap-ylococcus aureus is inducing many infections and it can infect tissues <-en t-e s1in and
mucosal +arriers' :-e plant Acant-us ilicifolius 093S -ave reported t-e follo<ing
p-ytoc-emicals, 28'2"Di =nor>c-olest5,",23trien22ol, 3met-o#ymet-o#y, (5purin8amine, N, (
+is =trimet-ylsilyl>?==trimet-ylsilyl> %#y>, 9yano colc-icines and 3+etamet-o#y5
c-oleston1(oic acid against a 12 essential protein and synt-etic drug of S' aureus <as collected
t-roug- literature survey' S' aureus <as studied @one of in-i+ition in different concentrations
against microorganisms and essential proteins <ere used as target <-ic- <ere doc1ed using
p-ytoc-emicals =ligands> found in A' ilicifolius leaves' In t-e present study, to analy@e t-e effect of
A' ilicifolius against S' aureus t-roug- doc1ing and in-i+ition studies, t-at A' ilicifolius -as
significant activity against S' aureus' :-is researc- <as -elpful attempt to t-e drug discovery in
S' aureus'
6ey <ords& Stap-ylococcus aureus, Acant-us ilicifolius, steroid components, target proteins
IN:;%D79:I%N
Acant-us ilicifolius /inn' =Acant-aceae>, locally 1no<n as Asea -ollyA is a spiny s-ru+ found
in t-e lo< lying coastal areas of sout-ern districts in 4anglades- and t-e vast area of mangrove
forest, t-e Sunder+ans and it is also <idely distri+uted' It is a viny s-ru+ or tall -er+ and +us-y
<it- very dense gro<t- =/a1s-mi et al', 1((">' 3any p-ytoc-emicals are present in A' ilicifolius
<-ic- is responsi+le for antimicro+ial, c-emo preventive, -epatoprotective, antiosteoporotic activity and
antiinflammatory activities =4a+u et al', 2001B Ags-i1ar et al', 1("(>' /ong +ac1, t-e leaf )uice of A'
ilicifolius <as commonly used +ot- internally and e#ternally for treating r-eumatism, <ounds,
neuropat-y and sna1e +ites' :-e seeds <ere ground and t-eir paste <as administered for +oils' In
Ayurvedic medical systems, A' ilicifolius is as an astringent, nervine tonic, stimulant and e#pectorant'
.arlier 093S studies <as done in our la+oratory s-o<n t-at t-e follo<ing ma)or
components present <ere in t-e A' ilicifolius leaves28,2"di =nor>c-olest5,",23trien22ol, 3
met-o#ymet-o#y, (5purin8amine, N, (+is =trimet-ylsilyl>?==trimet-ylsilyl> %#y>, 9yano
colc-icines and 3+etamet-o#y5c-olesten1(oic acid =4ala et al', 2011>'
Am' J' Drug Discov' Dev', 2013
S' aureus -as increased t-e ris1 of t-e infection development, +ot- nosocomial and
communityacCuired =De/eo et al', 2010B ;afee et al', 2012B Dang et al', 2010>' In 1(80
met-icillinresistant S' aureus' 3;SA, -as +ecome one of t-e diseaseproducing agent for
-ealt-care associated infections and itAs +ecome very difficult to treat =4igos and Denys, 200?>'
:-e present study aims to analy@e t-e p-ytoc-emicals +y 003S and evaluate t-e
antimicro+ial activity of most relevant compounds identified and Cuantified in A' ilicifolius'
Eurt-ermore, a Structureactivity ;elations-ip =SA;> analysis and molecular doc1ing studies
against S' aureus <ere performed, in order to provide t-e mec-anism of potential antimicro+ial
drugs for resistant microorganisms' 3olecular doc1ing is an in silico tool t-at predicts -o< a ligand
interacts <it- a receptor and -as +een successfully applied in several t-erapeutic programme at t-e
lead drug discovery =0-os- et al', 2008>'
3A:.;IA/S AND 3.:5%DS
Anti+acterial activity
Fone of in-i+ition& Acant-us ilicifolius <ere +otanically identified +y t-e 4otanical survey of
India, :amil Nadu Agriculture 7niversity, 9oim+atore' A vouc-er specimen of t-e plant -as +een
deposited at t-e +otanical survey of India -er+arium =2ouc-er num+er535?2>' Eres- leaves of
A' ilicifolius <ere cleaned <it- deioni@ed <ater and dried in t-e s-ade' :-e dried leaves <ere ground
and sieved' So#-let e#trtaction <as carried out for t-e Acant-us ilicifolius leaf samples <it-
solvent met-anol =%1unade et al', 200">'
3icroorganism <as gro<n overnig-t at 3"G9 in nutrient +rot- =,ere@ et al', 1((0>' :-e
antimicro+ial activity of t-e crude leaf e#tract of A' ilicifolius <as studied in different
concentrations against +acterial strain, t-e antimicro+ial potential of leaf e#tract <as assessed in
terms of @one of in-i+ition of +acterial gro<t-'
Anti+acterial activity
In silico analysis& :-e list of 12 genes involved in S' aureus infection pat-<ay in -uman <as
collected from literature =4artlett and 5ulten, 2010>' :-ese <ere found to +e essential for t-e
survival of t-e organism as reported in t-e D.0 data+ase =<<<' deg' %rg>' :-ey are Alp-a
-emolysin, ,anton 2alentine leu1ocidin, .nteroto#in, :o#ic s-oc1 syndrome to#in1,
Stap-ylo1inase, Aureolysin, 9lumping factor A, 9lumping factor 4, Sec A protein translocase,
Serineaspartate repeatcontaining protein 9, 30S ri+osomal protein S!, Serineaspartate
:a+le 1& 0enes and proteins of S' aureus responsi+le for infection
0enes ,roteins 7niprot ID
5la Alp-a -emolysin H"A832
/u1 ,anton valentine leu1ocidin
.nt0 .nteroto#in ,0A0/"
:st :o#ic s-oc1 syndrome to#in1 H"A!6"
Sa1 Stap-ylo1inase H((S7"
Aur Aureolysin H"A3"?
9lfA 9lumping factor A H((2J!
9lf4 9lumping factor 4 H"A3?2
Sec A Sec A protein translocase H"A8;5
Sdrc Serineaspartate repeatcontaining protein 9 H"A"?1'
Sas 0 30S ri+osomal protein S! ,88583
9an Serineaspartate repeatcontaining protein D ,88583
Am' J' Drug Discov' Dev', 2013
repeatcontaining protein D =:a+le 1>' :-ese proteins <ere ta1en as targets for doc1ing <it-
p-ytoc-emicals identified in A' ilicifolius' :-e structure of all target proteins for
Stap-ylococcus aureus <as retrieved from ,D4 =,rotein Data 4an1>' :-e +inding site of all target
proteins <as found using t-e H site finder for grid generation in Sc-rodinger' Identifying t-e
ligand +inding sites on a protein is t-e functional importance of many applications including
molecular doc1ing, de novo drug design and structural identification and comparison of functional
sites'
ISIS Dra< 2'3 soft<are <as used to design t-e p-ytoc-emicals' Analogues <ere c-anged in
3%/ files and 3D optimi@ation <as done +y 9-emS1etc- 3D vie<er of A0D/A4S ?'0' :-e 3%/ files
<ere converted into ,D4 format using 2.0 FF soft<are' :-e doc1ing studies -ave +een used <it-
Sc-rodinger 0lide program version !'0' :-e +est 10 poses and corresponding scores -ave +een evaluated
using 0lide in single precision mode =0lideS,> for eac- ligand' Eor eac- screened ligand, t-e pose <it-
t-e lo<est 0lide S, score -as +een ta1en as t-e input for t-e 0lide calculation in e#tra precision
mode =0lide I,>' :-e doc1ing <as carried out <it- 0lide S, and 0lide I,'
;.S7/:S AND DIS97SSI%N
A' ilicifolius met-anolic e#tracts -as s-o<n significantly -ig-er <-en compared to met-anolic
control =pJ0'05>' In order to verify t-e results, t-e e#periment <as triplicated and 3eanKSD <as
reported =:a+le 2>' :-e p-ytoc-emical reported in A' ilicifolius <ere considered as ligands for
doc1ing' :-ese essential proteins in S' aureus <ere used as target proteins <-ic- <ere doc1ed using
p-ytoc-emicals =ligands> found in Acant-us ilicifolius leaves using Sc-rodinger soft<are'
=:a+le 3>'
Alt-oug- t-e p-ytoc-emicals e#-i+ited +onding <it- proteins present in S' aureus, t-eir
interaction <as found to +e less <-en compared to common drug ,enicillin 0' So it is suggested t-at
alt-oug- t-e p-ytoc-emicals present in Acant-us ilicifolius leaves -as individual insignificant
activity against S' aureus =pJ0'05>, t-ere may +e a synergy effect of p-ytoc-emicals toget-er in
in-i+iting t-e gro<t- of S' aureus as seen in t-e @one of in-i+ition' :-is researc- <as an attempt to
ta1e a one step to<ards t-e drug discovery for Stap-ylococcus aureus'
:a+le 2& Anti+acterial activity of A' ilicifolius
9oncentration =mg m/ #> Fone of in-i+ition =mm>
80 13'333K2'0?
?0
13'333K2'0?
100
18'333K1'52
9ontrol =met-anol> "K0'5"00000
:a+le 3& Doc1ing of p-ytoc-emicals present in t-e leaves of A ilicifoilus <it- S' aureus target proteins
S' No' /igand ,roteins No' of +onds Doc1ing score 0lide score
3p met-o#y5c-olesten 1(oic acid Aur 3'82(00" 31'812585
9an 2'(5?888 23'"?53"1
9lfA 1'(501"8 35'(28"?!
.nt0 !'0?"833 31'8"!3"L
Sas 1'0525?0 3('20(318
clf4 1'!0830" 30'"2?!0!
(5purin8amine, N, (+is Aur 3'!0(015 !3'?118"0
=trimet-ylsilyl>?==trimet-ylsilyl> o#y> 9lfA 3'5!1831 !3'5?(035
.nt0
3'"?8?35 !0'382101
Sa1 2'"(2525 3?'802281
Am' J' Drug Discov' Dev', 2013
:a+le 3& 9ountinue
S' No' /igand ,roteins No' of +onds Doc1ing score 0lide score
Sas0 2'1?1815 !3'!2!218
clf4 3'513!(2 33'8"003"
28'2"Di =nor>c-olest5, ", Aur 1'"3""33 !0'03?("!
23trien22ol, 3met-o#ymet-o#y 9an 2'!2""!" 1('"!?5!5
9lfA !'0?0"?3 38'320?12
.nt0 3'"!8805 28'1530!"
Sa1 3'0!8!?( 32'!1"228
Sas0 2'5122(5 2('1?1025
9lf4 5'331(3" 30'8?0528
9yano colc-icines clf4 2'13""!8 3?'0(01!(
9lfA 2'30205! 28'?88""2
.nt0 2'1(?!"8 2?'!0351!
Sa1 1'?511"0 2"'?0(3?(
Sas0 1'??83(" 3('82"!1"
,enicillin 0 clfA !'2"(8(8 3"'5238!3
Sas0 2'?5(228 2('5"512L
Aur 5'"(130! "2'38(!5!
Sa1 2'"10(!? 2!'"53152
clf4
5'111300 !3'28210"
Sdr9 2'2?81?2 2('88??13
SecA 5'!(?11? 3!'(5?50(
/u1 5'1!02(5 3!'(5?50(
;.E.;.N9.S
Ags-i1ar, N'2', 2';' Nai1, 0J' A+ra-am, 9'2' ;eddy, S'D' NaCvo and ,'6' 3ittal, 1("('
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Am' J' Drug Discov' Dev', 2013
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