0.%*# #.'2%3!2%0!4-)')5*.'#+#.'2%3! 6$%!7859#%:);-
TRANSCRIPT
!
! !
!
"#$#%&$'()*#+,-.!/0.%*##.'2%3!2%0!4-)')5*.'#+#.'2%3!
6$%!7859#%:);-<%3#3.%!=#%)+.%!2%0!7859#%.%!!
0.+!>)')+#*.%!Eimeria tenella!2%0!0.'.%!?2*12%3!#(!!
')*#$%)9.%!@#'5+*$&&0.+#3%!!
!
!
!
A#++.'*)*#$%!
"#$!%$&'()#()!*+,!-$'*+,!
./0120$!*+$!3'2#$45,,+(,67'82+(.!
!
!
!
'9!:'67;+$+567!<50&0)5+!
*+$!=07'((+,!-#2+(;+$)>?(5@+$,52A2!
5(!B'5("!
!
!
!
!
!
='#+*#%!B%3.9+!
)+;0$+(!'9!CDEDFEGHIF!5(!B'5("!
!
!
!
!
B'5("J!CDGD!
!
!
!
!
!
!
!
!
!
!
!
!
I.#%.%!B9*.'%!
!
!
! !
!
!
!
!
!
!
!
!
!
!
!
Logic is blind and often knows only its own pastE!
>K$675@+,!8$09!-+(+256,!20!L75&0,0M7NJ!
609M5&+*!;N!27+!O0$6+$+,,+,!08!P0,,'1!
!
!
!
!
!
!
!
!
!
!
! !
Q$';.(.'52%3!!
:$+9*,M$'67&567+! K#,*$Q61+! #(*! :'672+$95(5J! 9+5,2! '#,! *+9! K()&0'9+$51'(5,67+(! ,5(*!
*#$67! 1#$,5@+! O67$+5;4+5,+! )+1+(("+567(+2E! /5+! R+$4+(*#()! ,0&67+$! K()&5"5,9+(! 10((2+!
(5672!599+$!@+$95+*+(!4+$*+(E!<+5!)A()5)+(!K#,*$Q61+(!45+!"#9!<+5,M5+&!SO0824'$+T!#(*!
SU'$*4'$+T!4#$*+!'#8)$#(*!*+$!;+,,+$+(!V+,;'$1+52!'#8!*5+!1#$,5@+!W+(("+567(#()!@+$"567>
2+2E!!
! INHALTSVERZEICHNIS
X!
/%-)9*+6.'1.#,-%#+!
H! B#%9.#*2%3 DDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDD H!GEG! U#9'(+>!#(*!25+$M'270)+(+!L'$',52+(!*+$!Apicomplexa EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE G!GEC! /+$!25+$M'270)+(+!L'$',52!Eimeria tenellaEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE Y!GEY! W01"5*50,+ EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE Z!GEYEG! W01"5*50,+!*+$!UQ7(+$EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE Z!GEYEC! [7+$'M5+!#(*!L$0M7N&'\+EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE I!GE]! W5(',+(!'&,!^5+&M$02+5(+!5(!*+$!_5$1,208880$,67#() EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE I!GEZ! ^N1&5(>';7A()5)+!W5(',+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEZEG! :#(1250(!*+$!^N1&5(>';7A()5)+(!W5(',+(!59!+#1'$N025,67+(!^+&&"N1&#,EEEEEEEEEEEEEEEEEE F!GEZEC! O2$#12#$+&&+$!K#8;'#!#(*!K125@5+$#()!@0(!^N1&5(>';7A()5)+(!W5(',+( EEEEEEEEEEEEEEEEEEE F!GEZEY! K[L><5(*+2',67+!@0(!^N1&5(>';7A()5)+(!W5(',+(!('67![$'\&+$EEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEZE]! K#8;'#!*+$!^N1&5(+EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEZEZ! ^N1&5(>';7A()5)+!W5(',+(!'&,!^5+&M$02+5(+!"#$!%(24561&#()!(+#+$!K(25M'$',5251'EEE F!GE`! <505(80$9'25,67+!K('&N,+!"#$!X*+(2585"5+$#()!@0(!^N1&5(>';7A()5)+(!W5(',+(EEEEEEE F!GEI! O2$#12#$;',5+$2+,!$'250('&+,!_5$1,2088*+,5)(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEF! Modelling!#(*!609M#2+$)+,2Q"2+,!_5$1,2088*+,5)( EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEFEG! U090&0)5+90*+&&5+$#()EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEFEC! B0&+1#&'$+,!DockingEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEFEY! O#;,2$#12#$>!#(*!a7(&5671+52,,#67+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEFE]! R5$2#+&&+,!Screening EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEFEZ! VX%>O59#&'250(+(!"#$!<+$+67(#()!*+$!8$+5+(!<5(*#(),+(+$)5+ EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEFE`! V+52,2$#12#$0M2595+$#()EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!GEH! K#8)';+(,2+&&#()EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!
J! I)*.'#)9!2%0!I.*-$0.% DDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDD R!CEG! U'$*4'$+!#(*!O0824'$+ EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEGEG! U'$*4'$+ EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEGEC! O0824'$+EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEC! /'2+(;'(1+( EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEY! B+270*+(!"#9!O+b#+("@+$)&+567EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEYEG! O+b#+(">Alignments EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CE]! X*+(2585"5+$#()!@0(!c/W>A7(&567+(!#(*!^N1&5(>A7(&567+(!L$02+5(+( EEEEEEEEEEEEEEEEEEEEEEE F!CEZ! U090&0)5+90*+&&5+$#()!*+$!%2BPW!#(*!*+$!L8BPW EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CE`! O#;,2$#12#$>!#(*!a7(&5671+52,,#67+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEI! R0$;+$+52#()!*+$!O#;,2'("*'2+(!8Q$!*',!90&+1#&'$+!/0615()EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!
INHALTSVERZEICHNIS
XX!
CEF! Docking EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!CEFEG! -dV/EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!CEFEC! LVK3[OEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!CEFEY! %5()+,+2"2+!<+4+$2#(),9+270*+( EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!CEFEYEG! PBO/>_+$2 EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEFEYEC! OM+'$9'(!W0$$+&'250(,10+885"5+(2!eP,f EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEFEYEY! K?c>_+$2 EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEFEYE]! K($+567+$#(),8'120$!e%:f EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!CEH! Docking *+$!+$7'&2+(+(!O#;,2'("+(!('67!O#;,2$#12#$,#67+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!CEGD! R5$2#+&&+,!Screening!('67!M02+(25+&&+(!%2BPW>!#(*!L8BPW>X(75;520$+(EEEEEEEEEEEEEEEEEF!CEGG! V5(+'$!X(2+$'6250(!%(+$)N!eVX%f!<+$+67(#()+( EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!CEGC! -+(+$5+$#()!+5(+$!109;5('20$5,67+(!O#;,2'("*'2+(;'(1 EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!CEGY! Vg,&5671+52,@0$7+$,')+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!CEG]! In vitro!B+,,#()+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!
P! B'3.;%#++.DDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDD R!YEG! <505(80$9'25,67+!O+b#+("'('&N,+!@0(!c/W,!#(*!^N1&5(+(!@0(!!E. tenellaEEEEEEEEEEEEEEF!YEGEG! c/W>7090&0)+!L$02+5(+!5(!E. tenella EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YEGEGEG! %2BPWEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!YEGEGEC! %2cPWGEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!YEGEGEY! %2cPWYEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!YEGEC! ^N1&5(+!@0(!E. tenella EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YEC! dM2595+$#()!#(*!K($+567+$#()!@0(!%2cPWC>X(75;520$+( EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YECEG! O#;,2$#12#$>!#(*!a7(&5671+52,,#67+( EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YECEC! B0&+1#&'$+,!Docking!"#$!K#,4'7&!@5$2#+&&+$!Hits EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YECEY! In vitro![+,2#()!*+$!'#,)+4A7&2+(!O#;,2'("+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YECE]! O2$#12#$>K125@52A2,><+"5+7#()+(!#(*!K('&N,+!*+$!L$02+5(>V5)'(*+(><5(*#()EEEEEEEEEF!YECEZ! R'&5*5+$#()!*+$!VX%>B+270*+!#(*!<+$+67(#()!*+$!-+45672#(),8'120$+(EEEEEEEEEEEEEEEEF!YECE`! <5(*+90*5>R0$7+$,')+!*+$!<+("595*'"0&>c'$;0(52$5&+ EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YECEI! In silico!dM2595+$#()!*+$!<+("595*'"0&>c'$;0(52$5&+ EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YEY! X*+(2585"5+$#()!@0(!%2BPW>!#(*!L8BPW>X(75;520$+( EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YEYEG! U090&0)5+90*+&&5+$#()!*+$!%2BPW!#(*!*+$!L8BPWEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YEYEC! %@'+$#()!*+$!Docking>O0824'$+!LVK3[OEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEF!YEYECEG! P+>Docking!%\M+$59+(2+EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!YEYECEC! K($+567+$#(),+\M+$59+(2+ EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!YEYEY! R5$2#+&&+,!Screeening!('67!M02+(25+&&+(!X(75;520$+(!*+$!%2BPW!#(*!*+$!L8BPW EEEEF!
!
! INHALTSVERZEICHNIS
XXX!
S! A#+52++#$%DDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDD R!]EG! ^N1&5(>';7A()5)+!W5(',+(!'&,!^5+&M$02+5(+!*+$![7+$'M5+!@0(!L$020"0+(>!
X(8+1250(+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]EC! <505(80$9'25,67+!X*+(2585"5+$#()!#(*!c7'$'12+$5,5+$#()!@0(!^N1&5(>';7A()5)(+(!
W5(',+(!5(!E. tenellaEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]EY! <505(80$9'25,67+!X*+(2585"5+$#()!#(*!c7'$'12+$5,5+$#()!@0(!^N1&5(+(!5(!!
E. tenella EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]E]! O2$'2+)5+(!"#$!U+99#()!@0(!W5(',+(EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]EZ! V+52,2#$12#$5*+(2585"5+$#()!@0(!X(75;520$+(!*+$!%2cPWC EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]E`! <5(*+90*5>R0$7+$,')+!#(*!dM2595+$#()!*+$!<+("595*'"0&+ EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]EI! U090&0)5+90*+&&5+$#()!*+$!%2BPW!#(*!*+$!L8BPW EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]EF! R'&5*5+$#()!*+$!O0824'$+!LVK3[O EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]EH! R5$2#+&&+,!Screening!('67!%2BPW>!#(*!L8BPW>X(75;520$+( EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!]EGD! K#,;&561EEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE F!
T! 72+)((.%&)++2%3 DDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDD R!
U! V#*.')*2'6.'1.#,-%#+ DDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDD R!
!
ABKÜRZUNGSVERZEICHNIS
XR!
W;5N'12%3+6.'1.#,-%#+!!Y/! ! ! /$+5*59+(,50('&!h! ! ! h(),2$g9!K<P! ! ! K*+(5(><5(*+$+)50(!K66E>3$E! ! Accession>3#99+$!K[L! ! ! K*+(0,5(2$5M70,M7'2!K?c! ! ! Area Under Curve!<;! ! ! Babesia bovis!<VKO[!! ! Basic Local Alignment Search Tool <VCO%i! ! Blast two Sequences!<VdO?B! ! Blocks Substitutions Matrix cKW! ! ! CDK-activating Kinase cKBW!! ! Ca2+/Calmodulin-dependent Protein Kinase!6*6! ! ! cell-division-cycle!c/W! ! ! Cyclin-dependent Kinase c7! ! ! Cryptosporidium hominis c7WG! ! ! Checkpoint Kinase 1!c5M! ! ! CD-interacting Protein c9! ! ! Cryptosporidium muris!cM! ! ! Cryptosporidium parvum cPW! ! ! CDK-related Kinase cN6! ! ! Cyclin /3K! ! ! Desoxyribonucleic Acid %:! ! ! Enrichment Factor!%-:>P!! ! Epidermal-Growth-Factor-Receptor!!%>value!! ! Expectation Value %O[! ! ! Expressed-Sequence-Tag %2! ! ! Eimeria tenella!%26N6! ! ! Eimeria tenella Cyclin -OWY! ! ! Glycogensynthase-Kinase-3!U,! ! ! Homo sapiens Xc! ! ! Inhibitor Concentration!Xc/! ! ! Intervet Chemical Database!X/! ! ! Identifier!X3W]! ! ! Inhibitor of CDK4 X(2+$L$0! ! Integrated Resource of Protein Domains and Functional Sites!1/'! ! ! W5&0*'&20(!BKL>W5(',+! ! Mitogen-activated Protein Kinase 3c<X! ! ! National Centre for Biotechnology Information BPW! ! ! MO15-related Kinase 3VO! ! ! Nuclear Localization Signal!LKB! ! ! Position Accepted Mutation L<P! ! ! L70,M7'2><5(*+$+)50(!L/<! ! ! Protein Data Bank LVL! ! ! Piecewise Linear Potential!L;! ! ! Plasmodium berghei!L66! ! ! Plasmodium chabaudi chabaudi!L8! ! ! Plasmodium falciparum
ABKÜRZUNGSVERZEICHNIS
R!
L8'9! ! ! Protein Domain Families L1! ! ! Plasmodium knowlesi!LW! ! ! L$02+5(!W5(',+!L@! ! ! Plasmodium vivax!LNN! ! ! Plasmodium yoelii yoelii!iOLP! ! ! Quantitative Structure Property Relationship P<P! ! ! P5;0,+><5(*+$+)50(!PX3-d!! ! Rapid Inducer of G2/M progression in Oocytes PBO/! ! ! Root Mean Square Deviation Pdc! ! ! Receiver Operating Characteristic
! ! EINLEITUNG
G!
H!B#%9.#*2%3!
!
HDH! X2()%.:!2%0!*#.'Y)*-$3.%.!>)')+#*.%!0.'!Apicomplexa!/5+!OM0$+(25+$67+(!0*+$!Sporozoen!eApicomplexaJ!,5+7+!K;;5&*#()!GEGf!,5(*!0;&5)'2+!%(*0>
M'$',52+(!@0(!_5$;+&&0,+(!#(*!_5$;+&25+$+(!eV?cX?O!k!VddO>:PK3WJ!CDDFfE!O5+!*#$67>
&'#8+(! +5(+(! 67'$'12+$5,25,67+(!-+(+$'250(,4+67,+&J! ;+5! *+9! ,567! )+,67&+672&567! #(*! #()+>
,67&+672&567!@+$9+7$+(*+!^+&&+(!';4+67,+&(!eB%UVUdP3J!CDDFfE!K9!R0$*+$M0&!*+$!@+)+>
2'25@+(! O2'*5+(! ;+,52"+(! *5+! OM0$0"0+(! +5(+(! ,M+"5'&5,5+$2+(! L&',25*+(! l! *+(! KM510M&',2+(E!
/5+,+$!5,2!('9+(,)+;+(*!#(*!67'$'12+$5,25,67!8Q$!*5+!-$#MM+!*+$!Apicomplexa!#(*!4+,+(2&567!
'9! X(@',50(,M$0"+,,! ,045+! *+$! %2';&5+$#()! *+,! L'$',52+(! 5(! *+$! _5$2,"+&&+! ;+2+5&5)2!
eB%UVUdP3J!CDDFfE!/+$!KM510M&',2!;+,52"2!+5(!+5)+(+,!L&',25*+(>-+(09!emYZ!1;fJ!4+&67+,!
*+9!M8&'("&567+$!L&',25*+(!A7(+&2!e:XcU%PK!k!PddOJ!GHHIfE!/5+,+,!-+(09!10*5+$2!(#$!6'E!
YD!L$02+5(+!e_XVOd3 et al.J!GHH`fE!:Q$!*5+!K#8$+672+$7'&2#()!*+$!;50,N(27+25,67+(!L$0"+,,+!
59!KM510M&',2+(!5,2!+,!*'7+$!(024+(*5)J!1+$(10*5+$2+!L$02+5(+!M0,22$'(,&'250('&!5(!*5+!L&',25*+!
"#!2$'(,M0$25+$+(E!B'(!)+72!*'@0(!'#,J!*',,!6'E!`DD!@+$,675+*+(+!L$02+5(+!l!65$6'!GD!L$0"+(2!
*+,!L'$',52+(M$02+09,!l!5(!*+(!KM510M&',2+(!2$'(,&0"5+$2!4+$*+(!9Q,,+(!e-KP/3%P et al.J!
CDDCn!^?%--% et al.J!CDDGfE!/5+
EINLEITUNG
C!
L'$',52! 1'((! @+$,675+*+(+!_5$2,0$)'(5,9+(J!45+! "E! <E!BA#,+! 0*+$! O674+5(+! ;+8'&&+(J! #(*!
+(24561+&2!*';+5!5(8+125g,+!#(*!45*+$,2'(*,8A75)+!^#,2'(*,80$9+(!e^N,2+(f!eBcKVVXO[%PJ!
CDDZfE!/+$!%(*45$2!5,2!5(!*+$!P+)+&!*5+!W'2"+J!*5+!*5+!5(8+125g,+(!%$$+)+$!*#$67!:$+,,+(!+5(+,!
5(85"5+$2+(! ^45,67+(45$2,! eB'#,f! '#8(5992! #(*! ('67! +5(+$! _+52+$+(24561&#()! *+$! [0>
\0M&',9+(J! 5(8+125g,+! d0"N,2+(! 952! *+9! W02! '#,,67+5*+2! eV?cX?O! k! VddO>:PK3WJ!
CDDFfE!K#8!*+(!B+(,67+(!Q;+$2$')+(!4+$*+(!*5+!%$$+)+$!Q;+$!*+(!+()+(!W0(2'12!"#!W'2"+(!
#(*!*5+!0$'&+!K#8('79+!*+$!d0"N,2+(!e"E!<E!W'2"+(102! 59!W'2"+(,2$+#fE!/5+![0\0M&',90,+!
@+$&A#82!;+5!*+(!9+5,2+(!B+(,67+(!,N9M209&0,!#(*!;+,674+$*+8$+5J!#(*!5,2!9+5,2!(#$!M$0;>
&+9'25,67!;+5!+5(+$!%$,25(8+1250(!@0(!O674'()+$+(J!*'!*+$!%$$+)+$!'#8!*',!?()+;0$+(+!Q;+$>
)+7+(!#(*!,674+$+!O67A*+(!5(!*+$!%9;$N0('&+(24561&#()!@+$#$,'67+(!1'((E![0\0M&',90,+!
1'((! '#67! 8Q$! X(*5@5*#+(!952! +5(+9! )+,2g$2+(! X99#(,N,2+9! )+8A7$&567! @+$&'#8+(J! *'! *5+,+!
1+5(+! '#,$+567+(*+! X99#(52A2! ;+,52"+(J! #9! +5(+! R+$9+7$#()! *+,! L'$',52+(! "#! @+$75(*+$(!
eBd3[djK!k!VX%O%3:%V/J!CDD]fE!!
!
Abbildung 1.1: Ausschnitt aus dem Kladogramm der Apicomplexa (MEHLHORN , 2001).
/+$! O2'99! *+$!Apicomplexa! #98',,2! 9+7$+$+! ;+*+#2+(*+! 7#9'(M'270)+(+! #(*! 25+$M'270)+(+! L'$',52+(! 45+!
;+5,M5+&,4+5,+!Eimeria sppEJ!Plasmodium sppEJ!Babesia sppE!#(*!Theileria sppE!!
Sporozoa (Apicomplexa)
Perkinsea Sporozoea
Gregarinia Coccidia
Agamococcidea EucoccideaProtococcidea
Eucoccida Haemosporoida
Adeleina Eimeriina Conoidina Aconoidina
Haemosporidae Piroplasmidae Klossiidae Eimeriidae
Klossia, Klossiella
Eimeria, Isospora,
Sarcocystis, Toxoplasma
Haemogregarinidae
Heptozoon, Haemogregarina,
Karyolysus
Plasmodium, Leucocytozoon, Haemoproteus
Babesia, Theileria
"#$%%!
&'$(()!
"*+,'$(()!
-+)./.01*12!
3.01*12!
41#)./.01*12!
5$%6'6)!
7$##*12!
! ! EINLEITUNG
Y!
/5+! /#$678'&&+$1'(1#()! c$NM20,M0$5*0,+! l! 7+$@0$)+$#8+(! *#$67! +5(+! X(8+1250(! 952! Cryp-
tosporidium parvum!l!5,2!;+5!)+,#(*+(!B+(,67+(!+;+(8'&&,!80&)+(&0,!#(*!7+5&2!5((+$7'&;!+5>
(+$!_067+!'#,!ecUKVB%PO!k!/KRX%OJ!CDDHfE!^#!*+(!4+52'#,!)+8A7$&567+$+(!%$1$'(1#(>
)+(!"A7&+(!B'&'$5'!%$1$'(1#()+(J!*5+!@0(!*+(!+5("+&&5)+(!L'$',52+(!*+$!-'22#()!Plasmodium!
7+$@0$)+$#8+(! 4+$*+(E! B'&'$5'! l! ;+1'((2! '#67! '&,! O#9M885+;+$! >! 5,2! +5(+! 9+&*+M8&56725)+!
W$'(17+52!#(*!5(!*+(!2$0M5,67+(!#(*!,#;2$0M5,67+(!VA(*+$(!4+52!@+$;$+52+2E!OM+"5+&&!;+2$088+(!
,5(*!4+52+
EINLEITUNG
]!
/+$!V+;+(,"N1&#,!@0(!E. tenella!5,2!5(!K;;5&*#()!GEY!,67+9'25,67!*'$)+,2+&&2E!^#(A67,2!4+$>
*+(! *5+! ,M0$#&5+$2+(! d0"N,2+(! @09!_5$2,0$)'(5,9#,! '#8)+(099+(E! /'('67! ,67&QM8+(! *5+!
OM0$0"052+(!59!/Q((*'$9!'#,!*+(!OM0$0"N,2+(!#(*!5(@'*5+$+(!*5+!%M527+&"+&&+(!*+,!/'$9,E!
%,! 10992! "#! +5(+$!9+7$1+$(5)+(! O675"0(2+(;5&*#()! 5(! +5(+$! M'$',520M70$+(!R'1#0&+E!K#,!
*+(! O675"0(2+(! )+7+(! ;+4+)&567+!B+$0"052+(! 7+$@0$J! 4+&67+! +5(+!4+52+$+! -+(+$'250(! @0(!
O675"0(2+(!5(!'(*+$+(!_5$2,"+&&+(!7+$@0$;$5()+(!1g((+(J!0*+$!"#!9A((&567+(!#(*!4+5;&567+(!
-'90(2+(!4+$*+(E!/5+!-'90(2+(!4+$*+(!'(,67&5+r+(*!"#!;+)+5r+&2+(J!9+7$1+$(5)+(!B51$0>
)'9+2+(! #(*! "#! +5(1+$(5)+(! B'1$0)'9+2+(! 4+52+$+(24561+&2E! B51$0>! #(*! B'1$0)'9+2+(!
@+$,679+&"+(!5(!*+(!%M527+&"+&&+(!"#$!^N)02+J!40$'#,!(+#+!d0"N,2+(!+(2,2+7+(J!*5+!952!*+9!
W02!*+,!_5$2,25+$+,!'#,)+,675+*+(!4+$*+(J!,0!*',,!*+$!^N1&#,!+$(+#2!;+)5((2E!K#r+$7'&;!*+,!
_5$2+,!85(*+2!*5+!OM0$#&'250(!*+$!d0"N,2+(J!;+5!707+$!V#828+#6725)1+52!#(*!K(4+,+(7+52!@0(!
O'#+$,2088J!5((+$7'&;!@0(!"4+5![')+(!,2'22E!%5(+!3+#5(8+1250(!*+,!_5$2,25+$+,!5,2!,0952!45+*+$!
9g)&567E!!!
!!
W;;#902%3!HDPC!V.;.%+18592+!6$%!Eimeria sppD![I.-9-$'%\!JOOH]D!
eGf!3'67!*+$!0$'&+(!K#8('79+!*+$!,M0$#&5+$2+(!d0"N,2+(!4+$*+(!*5+!OM0$0"052+(! 8$+5)+,+2"2!#(*!;+8'&&+(!*+(!
/Q((*'$9E!O5+!*#$67&'#8+(!*0$2!*5+!O675"0)0(5+!eC!;5,!Zf!,045+!*5+!-'90)0(5+!e`!;5,!GDf!;+@0$!,5+!'&,!#(,M0$#>
&5+$2+!d0"N,2+(!eGDf!952!*+9!W02!'#,)+,675+*+(!4+$*+(E!%,!80&)2!*',!O2'*5#9!*+$!OM0$0)0(5+!eGG!;5,!GYf!;+@0$!
,5+!@09!_5$2!45+*+$!'#8)+(099+(!4+$*+(!eGfE!
! ! EINLEITUNG
Z!
W;;#902%3! HDSC! "92*#3.! "9#%00<'(.! 0.+!
XN-%,-.%+!%),-!.#%.'!E. tenella!/%&.5*#$%!
K;;5&*#()!'#,!eB%UVUdP3 et al.J!GHHYfE!!
!
HDP! =$51#0#$+.!K&,!W01"5*50,+!;+"+567(+2!9'(!%$1$'(1#()+(!*#$67!%5("+&&+$J!*5+!*+$!W&',,+!*+$!Coccidia!
'()+7g$+(!eK;;5&*#()!GEGf!#(*!'&,!W01"5*5+(!;+"+567(+2!4+$*+(!eB%UVUdP3J!CDDFfE!/5+>
,+! %$1$'(1#()! "A7&2! "#! *+(! L$020"0+(5(8+1250(+(E! /5+! W01"5*5+(! ;+8'&&+(! @0$! '&&+9! *+(!
/'$9J!;+5!+5(5)+(
EINLEITUNG
`!
e,5+7+!K;;5&*#()!GE]f!!#(*!1'((!7A90$$7')5,67+!%(2+$525,!'#,&g,+(J!*5+!"#!+5(+$!707+(!O2+$;>
&5671+52,$'2+!p#()+$!Rg)+&!#(*!*'952!"#!707+(!g10(095,67+(!R+$&#,2+(!8Q7$2E!!!
Tabelle 1.1: Eimeria-Arten des Huhnes sowie Lokalisationen und pathologische Veränderungen (verändert
nach (KUMMERFELD, 2005)). !
Eimeria sppD!>'<Y)*.%1!
^M3_!V$5)9#+)*#$%! I$'*)9#*<*!
I)5'$+5$Y#+,-.!
V<+#$%.%!A2',-&)99!
E. tenella `! ^A1#9! ttt!UA90$$7')5,67+!
%(2+$525,!;)!
E. necatrix `!9522&+$+$!
/Q((*'$9!ttt!
UA90$$7')5,67+!
%(2+$525,!
,67&+595)J!
;)!
E. maxima Z!9522&+$+$!
/Q((*'$9!tt!
/'$9+$4+52+$#()J!
@+$*5612+!B#10,'J!
L+2+675+(!
,67&+595)J!
$0,'!
E. brunetti Z! X&+#9! tt!
W'2'$$7'&5,67>
7A90$$7')5,67+!!
%(2+$525,!
,67&+595)J!
;)J!
;$g61&5)!
E. acervulina ]!@0$*+$+$!
/Q((*'$9!t!
4+5r&567+!!
i#+$,2$+58#()!
6$+9+'$25)J!
,67&+595)!
E. mivati ]!@0$*+$+$!
/Q((*'$9!t! 4+5r+J!$#(*+!U+$*+!
6$+9+'$25)J!
,67&+595)!
E. mitis ]!@0$*+$+$!
/Q((*'$9!t! W+5(+! >!
E. praecox ]!@0$*+$+$!
/Q((*'$9!>! W+5(+! >!
E. hagani `!@0$*+$+$!
/Q((*'$9!>! W+5(+! >!
!
!
W01"5*50,+(!8Q7$+(!,M+"5+&&!5(!*+$!5(2+(,5@+(![5+$7'&2#()!"#!)$0r+(!L$0;&+9+(E!X(!*+$!UQ7>
(+$"#672!,5(*!4+&24+52!pA7$&567!$#(*!]D!B5&&5'$*+(!UQ7(+$!;+2$088+(!e/KVVd?V!k!VXVV%>
Ud=J!CDD`fE!/#$67!*5+,+!%$1$'(1#()!+(2,2+7+(!pA7$&567!4+&24+52!R+$&#,2+!5(!+5(+$!)+,67A2"2+(!
Ug7+!"45,67+(!ZDD!B5&&50(+(!u!eOUXPV%j et al.J!CDDIf!#(*!#(8',,;'$+(!Y!B5&&5'$*+(!?Ov!
e/KVVd?V!k!VXVV%Ud=J!CDD`fE!
! ! EINLEITUNG
I!
HDPDJ! M-.')Y#.!2%0!>'$Y-89)`.!K#67!;+5!+5(+$!,674+$+(!W01"5*50,++$1$'(1#()!45$*!9+5,2!(#$!,N9M209'25,67!27+$'M5+$2E!/5+!
,N9M209'25,67+![7+$'M5+!#98',,2!+5(+!'#,$+567+(*+!:&Q,,5)1+52,"#8#7$!>!(028'&&,!9#,,!,0)'$!
+5(+!<Q;+$2$')#()! +$80&)+(! >! #(*! *5+!-';+! @0(!%5,+(M$AM'$'2+(E!?(2+$,2Q2"+(*!45$*! *5+!
-';+! @0(! B#&25@52'95(M$AM'$'2+(! +9M807&+(E! -&+567"+525)! ,0&&2+(! '#67! *5+! UN)5+(+;+*5(>
)#()+(!@+$;+,,+$2!4+$*+(E!U5+$;+5!,M5+&2!*+$!K#,,67&#,,!@0(!8A1'&+$!W0(2'95('250(!*+,!:#2>
2+$,J!W0(2'12@+$9+5*#()!*+,![$5(14',,+$,!952! *+9!:+*+$1&+5*! *+$![5+$+J! #(*!O+(1#()!*+$!
V#828+#6725)1+52!+5(+!+(2,67+5*+(*+!P0&&+E!^#$!L$0M7N&'\+!*+$!W01"5*50,+!1g((+(!'#67!W01>
"5*50,2'251'!45+!O#&80('95*+J!/+60b#5('2J!K9M$0&5#9J![0&2$'"#$5&!#(*!/56&'"#$5&!+5()+,+2"2!
4+$*+(!eB%UVUdP3 et al.J!GHHYn!B%UVUdP3J!CDDFfE!/5+!+$,2+(!W01"5*50,2'251'!4#$*+(!
%(*+!*+$!YD5)+$!='7$+!+(24561+&2J!#(*!+$9g)&5672+(!*5+!W0(2$0&&+!*+$!W01"5*5+(M0M#&'250(+(J!
4',!+5(+!5(2+(,5@+!-+8&Q)+&7'&2#()!+$&'#;2+E!K#8)$#(*!7A#85)+$!P+,5,2+(";5&*#()!)+)+(Q;+$!
*5+,+(!_5$1,2088+(!5,2!*5+!%(24561&#()!(+#+$J!+88+125@+$!#(*!M$+5,4+$2+$!K&2+$('25@+(!p+*067!
*$5()+(*!+$80$*+$&567!eWX33KXP/ et al.J!CDD]n!B%UVUdP3!J!CDDGn!OUXPV%j et al.J!CDDIn!
O[%LU%3 et al.J!GHHIfE!/'$Q;+$!75('#,!+\5,25+$+(!'#67!'22+(#5+$2+!V+;+(*@'1"5(+!"#$!W01>
"5*50,+>L$0M7N&'\+J!45+!;+5,M5+&,4+5,+!L'$'60\w!e_XVVXKBO et al.J!GHHHfE!V+;+(*@'1"5(+>
$#()!5,2!"4'$!+88+125@J!p+*067!,+7$!2+#+$!#(*!g10(095,67!(5672!,5((@0&&!'#8!;$+52+$!%;+(+!5(!
*+$!5(2+(,5@+(!UQ7(+$"#672!+5(,+2";'$!eWX33KXP/ et al.J!CDD]n!Bc/d3KV/!k!OUXPV%jJ!
CDDHn!OUXPV%jJ!GHFHfE!
!
HDS! =#%)+.%!)9+!7#.9Y'$*.#%.!#%!0.'!@#'5+*$&&&$'+,-2%3!R0(!*+(!$#(*!CDEDDD!;5,!CZEDDD!-+(+(!*+,!9+(,67&567+(!-+(09,!eXU-OcJ!CDD]fJ!10*5+$+(!
ZGF!8Q$!L$02+5(15(',+(!e*5+,!+(2,M$5672!+24'!GJIq!'&&+$!9+(,67&567+(!-+(+fE!/'952!"A7&+(!,5+!
"#!+5(+$!*+$!)$gr2+(!L$02+5(8'95&5+(!eOU%X3%PBK3 et al.J!CDDZfE!L$02+5(15(',+(!8#()5+$+(!
'&,! L70,M70$N&2$'(,8+$',+(J! *E! 7E! ,5+! 2$'(,8+$5+$+(! +5(+! L70,M7'2)$#MM+! @0(! +5(+9! /0(0$!
eK[Lf! '#8! O+52+(1+22+(! ;+,25992+$! K95(0,A#$+(E! =+! ('67! OM+"5852A2! 4+$*+(! ,5+! 75+$;+5! 5(!
"4+5!-$#MM+(!+5()+2+5&2E!/5+!+5(+!-$#MM+!M70,M70$N&5+$2!L$02+5(+!'(!O+$5(>!#(*o0*+$
EINLEITUNG
F!
,67+(!4+$*+(!952!W$'(17+52+(! 5(!R+$;5(*#()!)+;$'672! eW3??[XVK et al.J!GHHFfJ!45+!;+5>
,M5+&,4+5,+!W$+;,+$1$'(1#()+(!eBKV?B<P%O!k!<KP<KcX/J!CDDHfJ!K*5M0,52',!e<KjOJ!
CDD]fJ
! ! EINLEITUNG
H!
Tabelle 1.2: Auf dem Europäischen Markt zugelassene Kinase-Inhibitoren (EMEA, 2009). !
@#'5+*$&&! @#'5$'*! X.'+*.99.'!a)-'!0.'!
729)++2%3!V#*.')*2'!
-&5@+6w!eX9'25(5;f!
8
8
8 89
89
3
88:9;
:9;
!
<6$>K;&! 30@'$25,! CDDG! eKjd?< et al.J!CDDZf!
X$+,,'w!e-+8525(5;f!
!"
#
$
$
$
%
%&
%
%
!
%-:P! K,2$'^+(+6'! CDD]!e3dPd!k!-%BBKJ!
CDDFf!
['$6+@'w!e%$&025(5;f!
8
83
3
33
:9;
:9;
89:9
!
%-:PJ!U%PG! P067+! CDDZ!e<KP%OcUX3d et al.J!
CDDIf!
3+\'@'$w!eO0$'8+(5;f!
8
389
3:9;
89
89
:':5;
3
!
6>P'8J!;>P'8J!
MYF'J!R%-:>
CJ!R%-:>YJ!
L/-: J!6>WX[!
<'N+$!
U+'&27c'$+!CDD`! eUK?OcUXV/J!CDDIf!
O#2+(2w!eO#(525(5;f!
89
35 8
9
89
3 8
:9;:9;
:9;
:9;
!
R%-:!G>YJ!
L/-: J!
L/-: J!6>WX[!
L85"+$! CDD`!ecUd?UK3 et al.J!
CDDIf!
OM$N6+&w!e/','25(5;f!
:9;
:'
89
3
"
8
89
88
88
39
:9;
!
<6$>K;&J!O$6J!
L/-: J!6>WX[!
<$5,20&>
BN+$,!
Ob#5;;!
CDD`! e<PKR% et al.J!CDDFf!
[',5)('w!e35&025(5;f!
8
8
89
8
:9;
89
3
8
8:9;
:5;
!
<6$>K;&J!
L/-:J!6>WX[!30@'$25,! CDDI!
ei?X3[KO>
cKP/KBK!k!cdP>
[%OJ!CDDIf!
EINLEITUNG
GD!
@#'5+*$&&! @#'5$'*! X.'+*.99.'!a)-'!0.'!
729)++2%3!V#*.')*2'!
[0$5,+&w!e[+9,5$0&59#,f!
$
$
$
$$&
$
$$ $$
$&$&
$
$&$
$
%&
&
& !
9[0$!
_N+27!
L7'$9'>
6+#256'&,!
CDDI!eBKVX^^XK!k!UO?J!
CDDFf!
[N@+$;w!eV'M'25(5;f!
8
83
89" 89
3
5
:'
3
3:9;
!
%-:PJ!U%PC!-&'\0>
O9527W&5(+!CDDF! x:PKBL[d3J!CDDHy!
!
!
!
<+5! *+$!%(24561&#()! @0(!L$02+5(15(',+>X(75;520$+(! ,5(*! M$5("5M5+&&! @5+$! @+$,675+*+(+!K$2+(!
*+$!X(75;5250(!*+(1;'$z!K[L>109M+2525@+!X(75;520$+(J!O#;,2$'2>109M+2525@+!X(75;520$+(J!X(75>
;520$+(! *+$! L$02+5(15(',+>K125@5+$#()! #(*! 5$$+@+$,5;&+! X(75;520$+(! xWV%<%J! CDDHyE! X(! *+$!
R+$)'()+(7+52! 7'2! 9'(! ,567! ('7+"#! '#,,67&5+r&567! '#8! +5(+! 109M+2525@+! R+$*$A()#()! *+,!
K[L,!@0(!,+5(+$!<5(*+,2+&&+!801#,,5+$2E!/5+,+!5,2!5((+$7'&;!*+$!L$02+5(15(',+(!)'("!;+,0(*+$,!
7067!10(,+$@5+$2J! ,0!*',,!,567!)#2+!O2$'2+)5+(!8Q$!*',!%5(M',,+(!@0(!X(75;520$+(!+(24561+&(!
&5+r+(E! K&&+$*5(),! 7'2! *5+! )$0r+! a7(&5671+52! *+$! K[L><5(*+,2+&&+(! *+(! 3'672+5&J! *',,! +,!
,6745+$5)!5,2J!+5(+(!X(75;520$!"#!+(24561+&(J!*+$!,+&+125@!*5+!<5(*+2',67+!+5(+$!)'("!;+,2599>
2+(!W5(',+!;&0615+$2E!/5+,!4#$*+!&'()+!^+52!'&,!#(9g)&567!+$'672+2E!/'$Q;+$!75('#,!,5(*!4567>
25)+!O5)('&2$'(,*#1250(,4+)+!7A#85)! $+*#(*'(2J!,0!*',,!*+$!K#,8'&&!+5(+,!_+)+,!Q;+$!+5(+(!
"4+52+(!_+)!109M+(,5+$2!4+$*+(!1'((!xWV%<%J!CDDHyE!%5(!+5("+&(+$!_+)!9#,,!,0952!(5672!
"4'(),&A#85)! +,,+(25+&&! ,+5(E! /5+! %(24561&#()! 109M+2525@+$! X(75;520$+(! *+$! K[L><5(*#()!
45$*! "#*+9!*'*#$67! +$,674+$2J! *',,! *+$!_5$1,2088!952! *+$! 59!95&&590&'$+(!<+$+567! 5(! *+$!
^+&&+!@0$7'(*+(+(!K[L>W0("+(2$'250(!x%-V%3!k!P%XOX3%J!CDDHy!10(1#$$5+$+(!9#,,J!#(*!
*5+! 9+5,2+(! L$02+5(15(',+(! W9>_+$2+! 8Q$! K[L! 59! 707+(! 951$090&'$+(! <+$+567! ;+,52"+(!
xWV%<%J!CDDHyE!/+$!+$,2+!K[L>W09M+2520$!X9'25(5;!e-&5@+6J!30@'$25,fJ!*+$!"#$
! ! EINLEITUNG
GG!
_5$1,2088! -+8525(5;! eX$+,,'wf! #(*! *+$! +\M+$59+(2+&&+! X(75;520$! -_>CDG`! x:K<XK3 et al.J!
CDDZyE!U+#2"#2')+!4+$*+(!5(,)+,'92!(+#(!X(75;520$+(!7#9'(+$!L$02+5(15(',+(!8Q$!@+$,675+>
*+(+!X(*51'250(+(!@+$9'$12+2!e,5+7+
EINLEITUNG
GC!
HDT! !7859#%:);-<%3#3.!=#%)+.%!!^N1&5(>';7A()5)+!W5(',+(!ec/W,!{!Cyclin-Dependent Kinasesf!;5&*+(!+5(+!-$#MM+!@0(!70>
90&0)+(J!7067!10(,+$@5+$2+(!L$02+5(15(',+(J!*5+!5(!'&&+(!%#1'$N0(2+(!"#!85(*+(!,5(*E!%5(5)+!
*5+,+$!L$02+5(+!,5(*!9'r)+;&567!'(!*+$!W0(2$0&&+!*+,!^+&&"N1&#,!;+2+5&5)2E!U5+$;+5!'125@5+$+(!
,5+! +5(+! R5+&"'7&! @0(! '(*+$+(! L$02+5(+(! *#$67! L70,M70$N&5+$#()J! *5+! "#! ;+,25992+(! ^+52>
M#(12+(!59!^+&&"N1&#,!;+(g25)2!4+$*+(E!c/W,!;5&*+(!W09M&+\+!952!*+(!"#)+7g$5)+(!^N1&5>
(+(J!40*#$67! ,5+! '&&0,2+$5,67! '125@5+$2!4+$*+(E!/5+!^N1&5(>';7A()5)+(!W5(',+(! ,5(*! (#$! 5(!
R+$;5(*#()!952! 57$+(!"#)+7g$5)+(!^N1&5(+(!'125@E!/'*#$67!1'((!*5+!K125@52A2!*+$!c/W,!5(!
*+$! ^+&&+! "+52&567! )+,2+#+$2! #(*! $+)#&5+$2! 4+$*+(E! /5+! ^N1&5(+! ,+&;,2! 4+$*+(! 75+$;+5! *#$67!
"+&&"N1&#,';7A()5)+!W0("+(2$'250(,A(*+$#()+(!$+)#&5+$2E!
!
HDTDH! b2%5*#$%!0.'!7859#%:);-<%3#3.%!=#%)+.%!#(!.25)'8$*#+,-.%!7.9918592+!/5+!^N1&5(>';7A()5)+(!W5(',+(!;+,52"+(!+5(+!456725)+!:#(1250(!5(!*+$!P+)#&'250(!*+,!+#1'>
$N025,67+(!^+&&"N1&#,E!/+$!+#1'$N025,67+!^+&&"N1&#,!;+,2+72!'#,!+5(+$!K;80&)+!7067!)+0$*(+>
2+$J!)&+567!';&'#8+(*+$!R+$A(*+$#()+(J!*5+!'&,!s;+$)A()+J!L7',+(!0*+$!'&,!O2'*5+(!;+"+567(+2!
4+$*+(E!U5+$"#!"A7&+(!*5+!GapG>L7',+!e-GfJ!*5+!/3K>ON(27+,+>L7',+!eOfJ!*5+!GapC>L7',+!
e-Cf! #(*! *5+! B520,+>L7',+! eBf! x%VV%/-%J! GHH`n! =dU3Od3! k! _KVW%PJ! GHHHy!
eK;;5&*#()!GEIfE!/+$!^+&&"N1&#,!;+)5((2!952!*+$!-G>L7',+E!X9!K(,67&#,,!80&)2!*5+!O>L7',+J!5(!
*+$!*5+!/3K!*+$!^+&&+!$+M&5"5+$2!45$*E!/'('67!4+$*+(!*5+!-C>L7',+!#(*!*5+!B>L7',+!*#$67>
&'#8+(E! /5+! +$,2+(! *$+5! L7',+(!4+$*+(! '#67! '&,! X(2+$M7',+! "#,'99+()+8',,2E!_A7$+(*! *+$!
B520,+!+$80&)2!*5+!+5)+(2&567+!^+&&2+5&#()J!#(*!*5+!K#82+5&#()!*+$!/3K!'#8!*5+!"4+5!(+#!+(2>
,2+7+(*+(
! ! EINLEITUNG
GY!
!!
Abbildung 1.7: Schema des eukaryotischen Zellzyklus und die Regulation durch Zykline sowie durch Zyklin-
abhängige Kinasen (nach [Malumbres&Barbacid,2009]).
/+$!^+&&"N1&#,!)&5+*+$2!,567!5(!*5+!;+5*+(!^45,67+(>L7',+(!-G!#(*!-CJ!*5+!ON(27+,+>L7',+!O!#(*!*5+!B520,+>
L7',+!BE!P+)#&5+$2!45$*!*5+,+$!^N1&#,!952!U5&8+!@0(!^N1&5(>';7A()5)+(!W5(',+(!,045+!*#$67!*+$+(!K125@'20$+(!
e^N1&5(+fE! /5+! c/W>^N1&5(>W09M&+\+! ,2+#+$(! *5+! s;+$)A()+! "45,67+(! *+(! +5("+&(+(! L7',+(E! /5+! W5(',+(!
c/WCJ!c/W]!,045+!c/W`!,5(*!+,,+("5+&&! 8Q$!*5+!L$0&58+$'250(!@0(! ,M+"5'&5,5+$2+(!^+&&+(! xBKV?B<P%O!k!
<KP<KcX/J!CDDHyE!!
X9!7#9'(+(!W5(09J!*',! '&&+!;+1'((2+(!#(*!9#29'r&567+(!W5(',+(!#98',,2J! ,5(*!GG!c/W!
L$02+5(+! ;+1'((2E! =+*067! ,5(*! (#$! +5(5)+! *+$! c/W! L$02+5(+! 5(! *+$! *5$+12+(! ^+&&"N1&#,>
P+)#&'250(! 5(@0&@5+$2! ec/WGJ! c/WCJ! c/W]! #(*! c/W`fE! /5+! c/W>L$02+5(+! ')5+$+(! '&,!
O67'&2+$J!*5+!*+(!s;+$)'()!@0(!*+$!-G!"#$!OG!L7',+!#(*!*+$!-C!"#$!B!L7',+J!,045+!'&&+!'(>
*+$+(!O67$522+! 5((+$7'&;!*+,!^+&&"N1&#,! ,2+#+$(E!O5+!;5&*+(!U+2+$0*59+$+!952! *+(!^N1&5(+(J!
40*#$67!,5+!'&&0,2+$5,67!'125@5+$2!4+$*+(E!<+59!B+(,67+(!,5(*!CH!^N1&5(+!;+1'((2J! p+*067!
')5+$+(! (#$! H! @0(! 57(+(! 59! ^+&&"N1&#,! e^N1&5(+z! KGJ! KCJ! <GJ! <CJ! /GJ! /CJ! /YJ! %GJ! %Cf!
xBKV?B<P%O! k! <KP<KcX/J! CDDZyE! /5+! ,0! '125@5+$2+(! W5(',+(! M70,M70$N&5+$+(!
[$'(,1$5M250(,$+M$+,,0$+(J! *5+! *'*#$67! 5('125@5+$2! 4+$*+(E! /5+! L70,M70$N&5+$#()! *#$67! *5+!
c/W>L$02+5(+! +$80&)2!9522+&,! *+,! L70,M7'2*0(0$,!K[L! #(*! 85(*+2! '(! O+$5(>! #(*![7$+0(5(>
$+,2+(!,2'22E!X(!*+$!:0&)+!45$*!*5+!%\M$+,,50(!@0(!-+(+(J!*5+!*5+!(A67,2+!L7',+!*+,!^+&&"N1&#,!
,2+#+$(J! 9g)&567E! W0(2$0&&5+$2! 4+$*+(! *5+! c/W,! *#$67! @+$,675+*+(+! :'120$+(z! "#9! +5(+(!
*#$67!*5+!<5(*#()!*+$!^N1&5(+J!"#9!'(*+$+(!';+$!'#67!*#$67!5(75;520$5,67+!?(2+$+5(7+52+(J!
45+!;+5,M5+&,4+5,+!*+$!c5M>!0*+$!X3W]>L$02+5(8'95&5+E!K#8!*5+,+!K$2!+(2,2+72!+5(!P+)+&1$+5,J!
*+$!*5+!,M+"585,67+(!L7',+(!*+,!^+&&"N1&#,!10(2$0&&5+$2!xBdP-K3J!GHHIyE!_5$*!*5+,+$!W$+5,>
&'#8J!"E<E!*#$67!,M+"585,67+!c/W>X(75;520$+(!#(2+$;$067+(J!5,2!*5+!^+&&@+$9+7$#()!)+7+992E!
<!7=!
"!
>!
>!
>!
!'()*
!+,"-.*/* !+,"-.*'*
!+,"-.*0* !+,"-.*1*
!'(2*345.67859-:,;8* <=8""8.*>.?*@5,9A97A4;8B*
!'(C*3/">9D-"?8.?8*=8""8.B*
!'(E*3(8-FG8""8.B*
7?!
EINLEITUNG
G]!
HDTDJ! G*'25*2'.99.'!W2&;)2!2%0!W5*#6#.'2%3!6$%!7859#%:);-<%3#3.%!=#%)+.%!/5+!:'&2#(),20M0&0)5+! *+$!L$02+5(15(',+(! 5,2! ,2'$1!10(,+$@5+$2! x<?X=OBK3J!CDD]yJ! #(*! ,5+!
"+5)+(! +5(+! 707+! O+b#+("A7(&5671+52! #(2+$+5('(*+$! x<PK3/%3! k! [dd^%J! GHHHyE! 3'67!
U'(1,!#(*!U#(2+$!xUK3WO!k!U?3[%PJ!GHHZy!&',,+(!,567!'&&+!L$02+5(15(',+>O2$#12#$+(!5(!
+&8!O#;*09A(+(!+5(2+5&+(!eK;;5&*#()!GEFfE!!!
!
Abbildung 1.8: Struktureller Aufbau von Proteinkinasen.
:'$;5)+!U+$@0$7+;#()!*+$!+&8!O#;*09A(+(!@0(!L$02+5(15(',+(!e('67!U'(1,!#(*!U#(2+$!xUK3WO!k!U?3[%PJ!
GHHZyf!'9!<+5,M5+&!^N1&5(>';7A()5)+$!W5(',+(J!L/<>X/z!G=O[E!!
K#67! 8Q$! *+(! c/WC>^N1&5(K>W09M&+\! ,5(*! *+2'5&&5+$2+! O2$#12#$5(80$9'250(+(! ;+1'((2!
x=%::P%j et al.J!GHHZyE!/+$!,2$#12#$+&&+!K#8;'#!*+$!c/WC!"+5)2!*',,+&;+!-$#(*)+$Q,2J!45+!
;+5! '(*+$+(! +#1'$N025,67+(! L$02+5(15(',+(E! /5+,+$! 4#$*+! "#9! +$,2+(! B'&! 8Q$! *5+! 6KBL>
';7A()5)+!L$02+5(15(',+! xW3X-U[d3 et al.J! GHHGy! 67'$'12+$5,5+$2E!/5+!c/WC>O2$#12#$! ;+>
=!?!
;!@!A!
B!
C!D!
=E!
F!
==!
!<H87F-.>:*
%<H87F-.>:*
I8"8.6<*78J-A.*3HingeB*
069-K-87>.J:<*:,;"8-L8*3H<LoopB*
! ! EINLEITUNG
GZ!
,2+72! '#,! +5(+$! 1&+5(+$+(! 3>2+$95('&+(! /09A(+! @0(! FZ! K95(0,A#$+(J! *5+! '#,! >:'&2;&'22>
O2$#12#$+(!'#8)+;'#2!5,2J!,045+!+5(+$!)$gr+$+(!c>2+$95('&+(!/09A(+E!O5+!#98',,2!CGY!K95>
(0,A#$+(!#(*!;+,52"2!+5(+(!707+(! >7+&51'&+(!K(2+5&!xBdP-K3J!GHHIn!LKRV%[XcUJ!GHHHyE!!!
!
!
Abbildung 1.9: Charakteristische Sequenzmotive von humanen CDK Proteinen.
^#! *+(! 67'$'12+$5,25,67+(! O+b#+("9025@+(! "A7&+(! LO[KXP%>B025@! e@50&+22fJ! ->Loop! e)+&;fJ! Hinge>P+)50(!
e)$Q(f!#(*!K125@5+$#(),>Loop!e$02fE!/',!^N1&5(><5(*+9025@!eLO[KXP%>B025@f!@'$55+$2!5(!*+$!O+b#+("';80&)+!5(!
7#9'(+(!c/W!L$02+5(+(E!/5+!K95(0,A#$+(!*+,!1'2'&N25,67+(!^+(2$#9,!WYCJ!%ZD!#(*!/G]]!e%2cPWC!3#99+>
$5+$#()f!,5(*!*#$67!S|T!9'$15+$2E!KG!;5,!K`!1+(("+567(+(!*5+!K[L><5(*#(),,2+&&+(E!/5+!L70,M70$N&5+$#(),,2+&>
&+(![G]J!jGZ!#(*![G`D!,5(*!+;+(8'&&,!;+5!7#9'(+(!c/W!L$02+5(+(!7067!10(,+$@5+$2E!!
!
c7'$'12+$5,25,67!8Q$!'&&+!c/W!L$02+5(+!5,2!+5(+!10(,+$@5+$2+!^N1&5(><5(*+*09A(+!eLO[KXP%J!
c/WCfJ!*5+! "#$! +5("5)+(! >U+&5\! ec>U+&5\f!*+$!3>2+$95('&+(!/09A(+!)+7g$2! #(*!+(2,67+5>
*+(*! 8Q$!*5+!^N1&5(;5(*#()! 5,2! e,5+7+!K;;5&*#()!GEHfE!K(!*+$!R+$;5(*#(),,2+&&+!*+$!;+5*+(!
/09A(+(! e'#67! '&,!Hinge>P+)50(! ;+"+567(+2! xBcX33%O! k! :XOcU%PJ! CDDZyfJ! &5+)2! +5(+!
25+8+!OM'&2+E!/0$2!;+85(*+2! ,567!*5+!7N*$0M70;+!K[L><5(*+2',67+E!/5+,+! 5,2J!+;+(,0!45+!*5+!
O#;,2$'2;5(*+2',67+J! 5(! *+$! 5('125@+(! :0$9! *#$67! +5(+! O67&+58+! e[>Loopf! ;&0615+$2E! /+$! [>
Loop! eK125@5+$#(),,67&+58+f! +(27A&2! *5+!K95(0,A#$+![7$G`D! e3#99+$5+$#()! ('67! 7#9'(+$!
c/WCfJ! *+$+(! L70,M70$N&5+$#()! "#! +5(+$! *$',25,67+(! K125@52A2,,2+5)+$#()! *+$! c/WC! 8Q7$2!
x=%::P%j et al.J! GHHZn!BdP-K3J! GHHIyE!/5+!<5(*#()! *+,! ^N1&5(,! '(! *5+! c/W! ;+45$12!
,2'$1+! R+$A(*+$#()+(! *+$! [+$25A$>O2$#12#$! *+$! L$02+5(+J! #(*! 5,2! +(2,67+5*+(*! 8Q$! *5+! O#;>
,2$'2,M+"5852A2E! U5+$;+5! 45$*! *5+! O#;,2$'2;5(*+,2+&&+! *+9',15+$2! #(*! *5+! 1'2'&N25,67+(! P+,2+!
eVN,YYJ!-&#ZG! #(*!K,MG]ZJ!3#99+$5+$#()! ('67! 7#9'(+$!c/WCf! 5(! )++5)(+2+$!_+5,+! 8Q$!
*+(!L70,M7'2)$#MM+(2$'(,8+$!@09!K[L!'#8!*',!O#;,2$'2!'()+0$*(+2E!/5+,!;+5(7'&2+2!*5+!<+>
4+)#()!*+,
EINLEITUNG
G`!
$#(),,67&+58+J!"45,67+(!5('125@+$!#(*!'125@+$!c/WC>O2$#12#$J!*+#2&567!"#!+$1+((+(!xUdVB
et al.J!CDDFn!3Xc?V%Oc?>/?RK^ et al.J!CDDIyE!!!
!
Abbildung 1.10: Vergleich der inaktiven und der aktiven CDK Struktur.
/+#2&567!"#!+$1+((+(!5,2!*5+!<+4+)#()!*+,!K125@5+$#(),>Loops!e$02!{!5('125@n!)$Q(!{!'125@f!,045+!*+$!c>U+&5\!
e)+&;fE!
!
HDTDP! WM>:"#%0.*)+,-.!6$%!7859#%:);-<%3#3.%!=#%)+.%!%),-!M')`9.'!X(! K(&+7(#()! '(! *',! K[L><5(*+2',67+(90*+&! ('67! [$'\&+$! x[PK}V%P et al.J! GHHHy! 8Q$!
K[L>109M+2525@+! W5(',+>X(75;520$+(! e,5+7+! K;;5&*#()! GEGGfJ! ;+,2+72! *5+! K[L><5(*+2',67+!
'#,!8Q(8!@+$,675+*+(+(!O#;2',67+(J!4+&67+!#(2+$,675+*&567+!M7N,51067+95,67+!%5)+(,67'82+(!
'#84+5,+(E!^#!*5+,+(!8Q(8!O#;2',67+(!"A7&+(!'f!*5+!K*+(5(><5(*+$+)50(!eK<PfJ!;f!*5+!P5;0>
,+;5(*+2',67+!eP<PfJ!6f!*5+!L70,M7'2><5(*+$+)50(!eL<PfJ!*f!*5+!<5(*+$+)50(!X!e<P>Xf!#(*!+f!
*5+! <5(*+$+)50(! XX! e<P>XXf! e,5+7+! K;;5&*#()! GEGGfE! K&&+! K[L>109M+2525@+(! W5(',+>
X(75;520$+(! ;5(*+(! 5(! *+$! 7N*$0M70;+(! K<PJ! #(*! 5(2+$')5+$+(! 952! *+$!Hinge>P+)50(! Q;+$!
_',,+$,2088;$Q61+(E!/5+!P<P!75()+)+(! 5,2! 7N*$0M75&! #(*!45$*! 082! *#$67! &g,&567+!-$#MM+(!
'*$+,,5+$2E! /5+,+! P+)50(! 5,2! 4+(5)+$! 10(,+$@5+$2! #(*! ;+5(7'&2+2! ,M+"585,67+! K95(0,A#$+(J!
4+&67+! 8Q$! +5(+! %$7g7#()! *+$! O+&+125@52A2! '#,)+(#2"2! 4+$*+(! 1g((+(! x<?X=OBK3J! CDD]n!
[PK}V%P et al.J! GHHHyE! /5+! [',67+! *+$! <P>X! ;+85(*+2! ,567! 5(! R+$&A()+$#()! *+,! 3`>
! ! EINLEITUNG
GI!
O2561,2088,!*+,!K[L,J!#(*!5,2!(5672!5(!*+$!<5(*#()!*+,!K[L,!5(@0&@5+$2E!/5+,+!P+)50(!5,2!(5672!
10(,+$@5+$2!#(*!45$*!)+(#2"2J!#9!,0407&!*5+!O+&+125@52A2!'&,!'#67!*5+!K125@52A2!@0(!X(75;520>
$+(!"#!+$7g7+(!x<?X=OBK3J!CDD]yE!/+$!^#)'()!"#!*5+,+$![',67+!45$*!*#$67!*5+!,0)+('((>
2+(! SGatekeeperT>K95(0,A#$+! 10(2$0&&5+$2! x<?X=OBK3J! CDD]n! -dPP% et al.J! CDDGyE! %5(!
^#)'()!"#!*5+,+$!<5(*+$+)50(!5,2!(#$!9g)&567J!4+((!'(!*5+,+$!O2+&&+!+5(+!,+7$!1&+5(+!K95(0>
,A#$+!eGatekeeperf!@0$&5+)2E!-+('#,0!45+!<P>X!5,2!'#67!<P>XX!(5672!'(!*+$!K[L><5(*#()!;+>
2+5&5)2E!/5+,+!O#;2',67+!1'((!+;+(8'&&,!"#!%$7g7#()!*+$!O+&+125@52A2!'&,!'#67!*+$!K125@52A2!@0(!
X(75;520$+(! )+(#2"2!4+$*+(! x<?X=OBK3J! CDD]yE!/5+! 7N*$0M70;+![',67+!L<P! 5,2! *+9!Vg>
,#(),9522+&!"#)+4'(*2!#(*!,67+5(2!8Q$!*5+!K885(52A2!@0(!X(75;520$+(!+5(+!+7+$!#(2+$)+0$*(+2+J!
'&&+(8'&&,!90*#&5+$+(*+!<+*+#2#()!"#!7';+(!x<?X=OBK3J!CDD]n!UdVB et al.J!CDDFyE!!!
!
Abbildung 1.11: ATP-Bindetasche nach Traxler.
/5+!<5(*+2',67+!&A,,2!,567!5(!Z!@+$,675+*+(+!O#;2',67+(!'#82+5&+(z!Kz!K<P!{!$02n!<z!P<P!{!@50&+22n!L<P!{!)+&;n!
<P>X!{!)$Q(n!<P>XX!{!;&'#E!!
!
HDTDS! W2&;)2!0.'!7859#%.!^N1&5(+!)+7g$+(!"#!+5(+$!,+7$!*5@+$,+(!L$02+5(8'95&5+E!X7$+!-$gr+!@'$55+$2!@0(!YZ!;5,!HD!1/'E!
K#8!*+$!M$59A$+(!O+b#+("+;+(+!,5(*!,5+!(#$!,+7$!)+$5()!10(,+$@5+$2E!/5+!*+((067!@0$7'(*+(+!
O+b#+("A7(&5671+52! 10("+(2$5+$2! ,567! '#8! +5(+(!K95(0,A#$+';,67(522J! ;+,2+7+(*! '#,! 6'E! GDD!
K95(0,A#$+(J! *+$! '&,! ^N1&5(><0\! ;+1'((2! 5,2E! /5+,+$! K;,67(522! 5,2! (024+(*5)! "#$! c/W>
<5(*#()!xWd<KjKOUX et al.J!GHHCn!V%%O!k!UKPVd_J!GHHYyE!K;)+,+7+(!@0(!*+9!)$0r+(!
O+b#+("#(2+$,675+*!;+,52"+(! p+*067!'&&+!^N1&5(+!+5(+!A7(&567+!-$#(*,2$#12#$E!O5+!#98',,+(!
Gatekeeper
EINLEITUNG
GF!
'&&+!59!W+$(;+$+567!GD!U+&56+,J!*5+!5(!C!U+&5\;Q(*+&(!'#,!p+4+5&,!Z!U+&56+,!eUG!l!UZ!#(*!UG~!
l!UZ~f!0$)'(5,5+$2!,5(*J!40;+5!*5+!^N1&5(><0\!*5+!+$,2+(!Z!U+&56+,!#98',,2!e,5+7+!K;;5&*#()!
GEGCfE!/5+,+!W+$(*09A(+!5,2!;+5,M5+&,4+5,+!;+5!^N1&5(!U!#(*!^N1&5(!K!;+,0(*+$,!,2'$1!10(>
,+$@5+$2E!/5+,!&+)2!*5+!R+$9#2#()!('7+J!*',,!*5+,+!^N1&5(+!+5(!$5)5*+,
! ! EINLEITUNG
GH!
HDTDT! 7859#%:);-<%3#3.!=#%)+.%!)9+!7#.9Y'$*.#%.!12'!B%*E#,592%3!%.2.'!W%*#Y)')+#*#5)!/5+!M$5("5M5+&&+!%5)(#()!@0(!c/W,!'&,!^5+&M$02+5(+! 8Q$!K(25M'$',5251'!4#$*+!;+$+52,! 5(!*+$!
V52+$'2#$! )+"+5)2! x/d%PX-J! CDD]n! UKOOK3 et al.J! CDDGn! Bd[[PKB et al.J! GHH`yE! /5+!
R+$9+7$#()!*+$!L$020"0+(!Plasmodium falciparum,!Leishmania mexicana!#(*!Trypanosoma
brucei!10((2+!*#$67!c/W>X(75;520$+(J!*5+!8Q$!7#9'(+!c/W,!+(24561+&2!4#$*+(J!#(2+$;#(*+(!
4+$*+(!x-PK3[ et al.J!CDD]n!UKPBO% et al.J!CDDGn!_dd/KP/ et al.J!CDDYyE!R0$@+$,#67+!
952!,0&67+(!X(75;520$+(!'(!E. tenella!7';+(!"#!A7(&567+(!%$)+;(5,,+(!)+8Q7$2!x<%j%PJ!CDDIyE!
U5+$"#!4#$*+!*+$!;+1'((2+!c/W>X(75;520$!:&'@0M5$5*0&! x^UKX et al.J! CDDCy! 5(! +5(+9!O675>
"0(2+(>P+58#(),>Assay!)+2+,2+2E!/5+,+!B+,,#()+(!4#$*+(!@0(!=0'6759!U089'((J!K(p'!U+>
61+$027!#(*!<$522'!@0(!d+M+(!eX(2+$@+2!X((0@'250(!-9;UJ!O674';+(7+59f!*#$67)+8Q7$2E!/5+!
O675"0(2+(>P+58#()!@0(!E. tenella!10((2+!952!:&'@0M5$5*0&>W0("+(2$'250(+(!@0(!GZD!(B!#(*!
YDD!(B!5(75;5+$2!4+$*+(E!X(75;520$10("+(2$'250(+(!#(2+$!FD!(B!"+5)2+(!1+5(+!_5$1#()E!W0(>
"+(2$'250(+(!0;+$7'&;!`DD!(B!4'$+(!p+*067!)5825)!8Q$!*5+!_5$2,"+&&+E!/5+!8Q$!:&'@0M5$5*0&!;+5!
+5(+$! W0("+(2$'250(! @0(! GZD! (B! 8+,2)+,2+&&2+! @0&&,2A(*5)+! X(75;5+$#()! *+$! O675"0(2+(>
P+58#()J!10$$+&5+$2!,+7$!)#2!952!*+$!X(75;5+$#(),10(,2'(2+!@0(!GG!(B!59!%("N9>Assay!+5(+$!
^N1&5(>';7A()5)+(!W5(',+!@0(!E. tenella! e%2cPWCfE!/+$!%("N9>Assay!4#$*+!@0(!B'$�'!VE!
O#Ä$+"!:+$(Ä(*+"!eX(2+$@+2!X((0@'250(!-9;UJ!O674';+(7+59f!*#$67)+8Q7$2E!
Tabelle 1.3: Ergebnisse der chemischen Validierung mit dem bekannten CDK-Inhibitor Flavopiridol. !
:&'@0M5$5*0&!x(By!
%2cPWC! U,c/WC!
XcZD! W5! XcZD! W5!%("N9+>Assay!
YY!Å!GD! GG!Å!Y! Y`!Å!GD! GH!Å!Y!
! ! ! ! !
O675"0(2+(!%(24561&#()!
GDDq!X(75;5+$#()! 1+5(+!X(75;5+$#()! ^+&&>[0\5"52A2!
GZD!
O675"0(2+(>
P+58#(),>Assay!
YDD!ÇFD! É`DD!
/'2+(!@0(!B'$�'!VE!O#Ä$+"!:+$(Ä(*+"! eX(2+$@+2! X((0@'250(!-9;UJ!O674';+(7+59n!%("N9>Assayf! ,045+! =0'>
6759!U089'((J!K(p'!U+61+$027!#(*!<$522'!@0(!d+M+(!eX(2+$@+2!X((0@'250(!-9;UJ!O674';+(7+59n!O675"0(2+(>
$+58#(),2+,2f!x#(M#;&5"5+$2+!/'2+(yE!!K("'7&!*+$!/#$678Q7$#()+(!59!%("N9+>Assayz!(!{!Y>]!,045+!59!O675"0(>
2+(>P+58#(),>Assayz!(!{!CE!
EINLEITUNG
CD!
HDU! "#$#%&$'()*#+,-.!W%)98+.!12'!/0.%*##.'2%3!6$%!7859#%:);-<%3#3.%!=#%)+.%!
?(2+$! 7090&0)+(! L$02+5(+(! @+$,2+72!9'(! L$02+5(+J! *5+! @0(! +5(+9! )+9+5(,'9+(!R0$8'7$+(!
';,2'99+(J!+)'&!0;!*',!B+$19'&!@+$A(*+$2!0*+$!#(@+$A(*+$2!@0(!*+9
! ! EINLEITUNG
CG!
&#()! +5(+,! 0M2595+$2+(! _5$1,2088,E! K$"(+59522+&J! *5+! #(2+$! 9'r)+;&567+$! <+2+5&5)#()! *+,!
_5$1,2088*+,5)(,!+(24561+&2!4#$*+(J!10((2+(!;+$+52,!+$80&)$+567! 59!B'$12!M&'2"5+$2!4+$*+(E!
3+#+$+!M$095(+(2+!<+5,M5+&+!*'8Q$!,5(*!-&5@+6w!xKjd?< et al.J!CDDZyJ!+5(!B+*51'9+(2!"#$!
<+7'(*&#()! *+$!c7$0(5,67+(!BN+&05*+(!V+#1A95+J!-&'@#,w! "#$!<+7'(*&#()! @0(!Diabetes
Mellitus![NM!C!xOcU%%3!k!LKi?d[J!CDDHyJ!['958&#w!#(*!P+&+("'w!xBKc/d3KV/J!CDDDy!
"#$! <+7'(*&#()! @0(! X(8&#+("'J! ,045+! K)+(+$',+w! ! x<PdLUj et al.J! CDDDyJ! X(@5$',+w!
xP%-K^^X et al.J!GHHHy!#(*!30$@5$w!xKPRX%?}!k![PX<?[J!CDDZyJ!*$+5!UXR!L$02+',+!X(75>
;520$+(!e,5+7+![';+&&+!GE]fE!!!
/',!,2$#12#$;',5+$2+J!$'250('&+!_5$1,2088*+,5)(!;+5(7'&2+2!59!_+,+(2&567+(!*5+!B+270*+(!@5$>
2#+&&+,! Screening! xWX[cU%3 et al.J! CDD]yJ! 90&+1#&'$+! V+52,2$#12#$0M2595+$#()! x/%K3J!
GHH]y! #(*! *',!de novo!/+,5)(! xOcU3%X/%P!k!:%cU3%PJ! CDDZyE!-$#(*&')+! *5+,+$! @+$>
,675+*+(+(!K,M+12+!*+,!$'250('&+(!_5$1,2088*+,5)(,!5,2!,2+2,!*5+!R0$7+$,')+!*+$!K885(52A2!*+,!
V5)'(*+(!"#9!+(2,M$+67+(*+(!P+"+M20$E!/5+!K885(52A2,@0$7+$,')+!;+$#72!'#8!*+$!<+4+$2#()!
*+$!90&+1#&'$+(!_+67,+&45$1#()+(!"45,67+(!*+9!L$02+5(J!*E7E!*+$!K95(0,A#$+(!59!'125@+(!
^+(2$#9!952!*+9!)+;#(*+(+(!V5)'(*+(E!<+5!*5+,+$!R0$7+$,')+!;+*5+(2!9'(!,567!,2$#12#$+&>
&+$J!20M0&0)5,67+$!#(*!M7N,51'&5,67>67+95,67+$!B+$19'&+J!*5+!9522+&,!609M#2+$;',5+$2+(!B+>
270*+(!;+$+67(+2!4+$*+(!1g((+(E!!
Tabelle 1.4: Beispiele für einige Medikamente die unter Beteiligung von rationalem Wirkstoffdesign entwickelt
wurden. !
@#'5+*$&&! 7#.9Y'$*.#%! X.'+*.99.'! =')%5-.#*!729)++2%3!
/%!B2'$Y)!
30$@5$w!eP520('@5$f!
! cN2067$09!
L]ZD!cjL!YK]!
K;;022!-9;U!
k!c0E!W-! UXR! GHH`!
P+&+("'w!e^'('95@5$f!
!
!
R5$'&+!
!3+#$'95(5*',+!
-&'\0O9527>
W&5(+!-9;U!k!
c0E!W-!X(8&#+("'! GHHH!
X(@5$',+w!eO'b#5('@5$f!
!cN2067$09>
L]ZD>
%("N9,N,2+9!
U0889'((>!
V'!P067+!K-! UXR! GHH`!
EINLEITUNG
CC!
@#'5+*$&&! 7#.9Y'$*.#%! X.'+*.99.'! =')%5-.#*!729)++2%3!
/%!B2'$Y)!
['958&#w!ed,+&2'95@5$f!
!R5$'&+!
!3+#$'95(5*',+!
U0889'((>!
V'!P067+!K-! X(8&#+("'! CDDC!
-&5@+6w!
!
K;+&,0(![N$0>
,5(>15(',+!
eK<Vf!
30@'$25,!K-!
c7$0(5,67+(!!
BN+&05*+(!!
V+#1A95+!
CDDG!
K)+(+$',+w!eK9M$+('@5$f!
!R5$'&+!L$02+',+!
UXR>G!
-&'\0O9527>
W&5(+!-9;U!k!
c0E!W-!
UXR! CDDD!
-&'@#,w!eR5&*')&5M25(f!
!
/5M+M25*N&>
L+M25*',+>]!
eDPP4f!
30@'$25,!K-!
/5';+2+,!!
9+&&52#,!!
[NM!C!
CDDI!
!
HDR! Modelling!2%0!,$(Y2*.'3.+*N1*.+!@#'5+*$&&0.+#3%!/',! $'250('&+! _5$1,2088*+,5)(! 45$*! 7+#2"#2')+! *#$67! &+5,2#(),8A75)+! c09M#2+$! #(2+$,2Q2"2!
x/UKVX_KV!k!cU%3J!CDDHyE!/5+,+!,5(*!5(!*+$!V')+J!%5)+(,67'82+(!@0(!B0&+1Q&+(!"#!90>
*+&&5+$+(E!X(!*+(!&+2"2+(!='7$+(!7'2!*',!Molecular ModellingJ!;+,0(*+$,!*',!B0*+&&5+$+(!*+$!
V5)'(*>P+"+M20$>_+67,+&45$1#()J! 5(! *+$! K$"(+59522+&80$,67#()! "#(+79+(*! '(! <+*+#2#()!
)+40((+(!x=XK3-J!CDDFyE!/5+!+5)+(2&567+!90&+1#&'$+!B0*+&&5+$#()!*5+(2!75+$;+5!@0$45+)+(*!
"#$! ,2$#12#$+&&+(!B0*5851'250(!@0(!V+52,2$#12#$+(J!4A7$+(*!;+59!,2$#12#$>!#(*!609M#2+$)+>
,2Q2"2+(!/+,5)(!,0407&!(+#+!V+52,2$#12#$+(!5*+(2585"5+$2!'&,!'#67!90*585"5+$2!4+$*+(!1g((+(!
x<KPPXV!k! OdVXRKJ! CDD`n! [K:[ et al.J! CDDFyE! 3+;+(! *+9!Modelling! 2$')+(! @0$! '&&+9!
b#'(252'25@+! O2$#12#$45$1#(),;+"5+7#()+(! "#9! R+$,2A(*(5,! *+$! ^#,'99+(7A()+! "45,67+(!
*+$!67+95,67+(!O2$#12#$!#(*!57$+$!_5$1#()!;+5!xcVKPWJ!CDDHyE!X(!*+(!80&)+(*+(!W'M52+&(!
,0&&! (#(! (A7+$! '#8! *5+!B+270*+(! *+,!Modellings! #(*! 609M#2+$)+,2Q2"2+(!_5$1,2088*+,5)(,!
+5()+)'()+(!4+$*+(E!X(!K;;5&*#()!GEGY!5,2!+5(!2NM5,67+$!L$0"+,,!*+$!V+52,2$#12#$5*+(2585"5+>
$#()!*'$)+,2+&&2E!!
! ! EINLEITUNG
CY!
!
Abbildung 1.13: Workflow zur Identifizierung potentieller Leitstrukturen.
/'$)+,2+&&2! ,5(*!*5+! ,+67,!U'#M2,67$522+! 59!V+52,2$#12#$5*+(2585"5+$#(),>L$0"+,,E!/5+,+$!L$0"+,,! 5,2! +5(!109;5>
(5+$2+$!K(,'2"!'#,!in silico!,045+!in vitro!K$;+52,67$522+(E!!!
HDRDH! X$($9$3#.($0.99#.'2%3!!-$#(*&')+! @5+&+$! B+270*+(! *+,! $'250('&+(!_5$1,2088*+,5)(,J! 45+! ;+5,M5+&,4+5,+! *+,! 90&+>
1#&'$+(!DockingsJ!5,2!*5+!W+((2(5,!*+$!*$+5*59+(,50('&+(J!90&+1#&'$+(!O2$#12#$!eY/>O2$#12#$f!
*+,!^5+&M$02+5(,!x<KPPXV!k!OdVXRKJ!CDD`yE!d829'&,!5,2!*5+,+!O2$#12#$!p+*067!(5672!;+1'((2!
#(*!1'((J!'#8)$#(*!*+$!K#84+(*5)1+52!*+,!+\M+$59+(2+&&+(!R+$8'7$+(,J!"#$!K;&+52#()!90&+>
1#&'$+$!Y/>O2$#12#$+(!;50&0)5,67+$!B'1$090&+1Q&+J!(5672!+5(8'67!;+,25992!4+$*+(!x[K:[ et
al.J!CDDFyE!%5(+!Bg)&5671+52J!*5+,+!B+270*+(!*+((067!'#67!'#8!^5+&M$02+5(+!#(;+1'((2+$!Y/>
O2$#12#$!'("#4+(*+(J!;5+2+2!*',!Homology Protein Modelling!xcKRKOd[[d!k!LUK[KWJ!
CDDHyE!-$#(*&')+!*5+,+,!R+$8'7$+(,!5,2!*5+!K(('79+J!*',,!7090&0)+!L$02+5(+!+5(+$!L$02+5(>
8'95&5+J!*5+!A7(&567+!K95(0,A#$+,+b#+("+(!'#84+5,+(J!Q;&567+$4+5,+!'#67!+5(+!A7(&567+!L$0>
2+5(8'&2#()!"+5)+(E!X,2!*5+!90&+1#&'$+!Y/>O2$#12#$!+5(+,!0*+$!9+7$+$+$!R+$2$+2+$!+5(+$!L$02+5(>
8'95&5+!;+1'((2J!1g((+(!*5+,+!P+8+$+(",2$#12#$+(!eTemplatef!'&,!O67';&0(+!8Q$!*5+!K;&+52#()!
+5(+,!90&+1#&'$+(!O2$#12#$>B0*+&&,!+5(+,!7090&0)+(!L$02+5(,!#(;+1'((2+$!O2$#12#$!)+(#2"2!
4+$*+(!x}XK3-J!CDD`yE!!!
R6.#*)'')(!Screening!
G.6%S.).!!in vitro Screen
T6/61U/.%$#6(VW)!I1$'X()1!
Y6)'O./#)61!J0)1#6U6Z6)M.*12!*10!R$'606).*12!!
In vitro!Hit !T)(#S#62*12!
GW$.%$V/OW/.)!56'#).*12!
:W)%6(VW)!!R$'606).*12!!
Docking
I*([$W'!0).!!R).+610*12)1!!
9/%/'/26)!!</0)''6).*12!!
8$VW+).)6#*12!!
N62$10)1!!R/.+).)6#*12!
J:AE!!T)(#6%%*12!
"#.*,#*.$1$'X()!!
G.6%S.).! in vitro Screen!!
In vitro Hit T)(#S#62*12!
J:AE!!T)(#6%%*12!
J1!\6#./!N)6#(#.*,#*.!T)(#S#62*12!
N:P<"!
8<K!
N]('6VW,)6#(M%)((*12)1!!
&6!T)(#6%%*12!
Hit!I1.)6VW).*12!
"*+(#.*,#*.M!*10!^W1'6VW,)6#(M(*VW)1!!
Docking
8$VW+).)6#*12!!
I*([$W'!0).!R).+610*12)1!!
N62$10)1!!R/.+).)6#*12!!
EINLEITUNG
C]!
!
Abbildung 1.14: Überlagerung der Homologiemodelle der Trehalose-6-Phosphat-Synthase (T6PS) von Dro-
sophila melanogaster und Ctenocephalides felis mit der Escherichia coli T6PS Kristallstruktur
1UQT.
Kz!d;+$8&A67+(*'$,2+&&#()!*+,!D. melanogaster!U090&0)5+90*+&&,!e$02f!#(*!*+$!E. coli W$5,2'&&,2$#12#$!G?i[!
e)+&;fE!<z!Ribbon>/'$,2+&&#()!*+,!C. felis!U090&0)5+90*+&&,!e$02f!#(*!*+$!E. coli!W$5,2'&&,2$#12#$!G?i[!e)+&;fE!
^#9! R+$)&+567! 4#$*+(! *5+! 9522+&,! B0&6'*! x<PXcWBK33J! GHHZn! <PXcWBK33! et alEJ! CDDDy! ;+$+67(+2+(!
d;+$8&A67+(!Q;+$+5('(*+$!)+&+)2E!!!
K9! K(8'()! *+$! U090&0)5+90*+&&5+$#()! ,2+72! "#(A67,2! *',! Alignment! *+$! K95(0>
,A#$+,+b#+("+(! *+,! L$02+5(,! 952! #(;+1'((2+$! O2$#12#$J! #(*! +5(+,! L$02+5(,! 952! ;+1'((2+$!
O2$#12#$!xi? et al.J!CDDHyE!=+!7g7+$!*5+!O+b#+("5*+(252A2!75+$;+5!5,2J!*+,20!;+,,+$!,5(*!'#67!*5+!
;+$+67(+2+(!U090&0)5+90*+&&+!x}XK3-J!CDD`yE!%(27A&2!*',!Alignment!)$gr+$+!VQ61+(;+$+5>
67+J!4+$*+(!K;,67(522+!*+,!;+1'((2+(!L$02+5(,! 5(!*',!B0*+&&! +5()+8Q)2E!<+5!(5672!Q;+$+5(>
,2599+(*+(!K95(0,A#$+(!'(!*+(!+5("+&(+(!L0,5250(+(!45$*!@+$,#672J!*5+!W0(80$9'250(,45(>
1+&!*+$!O+52+(1+22+(! +(2,M$+67+(*!*+$!d$)5('&,2$#12#$!;+5"#;+7'&2+(E!/5+,+!4+(5)+$!10(,+$>
@5+$2+(!<+$+567+!;5&*+(!082!O67&+58+(!eLoopsf!#(*!85(*+(!,567!5(2+$+,,'(2+$4+5,+!9+5,2!'(!*+$!
d;+$8&A67+!*+,!L$02+5(,E!O5+!"+5)+(!+5(+(!@+$)&+567,4+5,+!)+$5()+(!%5(8&#,,!'#8!*5+!,2$#12#>
$+&&!10(,+$@5+$2+(!P+)50(+(!59!X((+$(!*+,!L$02+5(,J!40!9+5,2!'#67!*',!'125@+!^+(2$#9!&01'&5>
,5+$2!5,2!xO%V^%P et al.J!CDDFyE!X9!K(,67&#,,!+$80&)2!+5(+!<+4+$2#()!*+,!B0*+&&,J!*5+!W0&&5>
,50(+(!"45,67+(!K209+(!;+$Q61,56725)2E!%@+(2#+&&!'#82$+2+(*+!K20910&&5,50(+(!4+$*+(!9'>
(#+&&!+(28+$(2!x<ÑUB et al.J!CDDCn!:dPO[%PJ!CDDCn!O%V^%P et al.J!CDDFyE!K#8!*5+,+!_+5,+!
+(2,2+72!+5(!L$02+5(90*+&&!*+$!O+b#+("J!"#!*+$!1+5(+!+\M+$59+(2+&&+(!O2$#12#$*'2+(!@0$&5+)+(!
e,5+7+!K;;5&*#()!GEG]fE!!
I! T
! ! EINLEITUNG
CZ!
HDRDJ! I$9.529)'.+!Docking!
K&,!Docking!;+"+567(+2!9'(!*',!609M#2+$)+,2Q2"2+!%5(M',,+(!1&+5(+$!67+95,67+$!O2$#12#$+(!
5(!*5+!<5(*+2',67+!+5(+$!90&+1#&'$+(!Y/>L$02+5(,2$#12#$!x<ÑUB et al.J!CDDCn!WV%<%J!CDDHn!
WPd%B%PJ!CDDIyE!/',!^5+&! 5,2!*';+5!*',!K#885(*+(!@0(!<5(*+90*5!9g)&567,2!)+$5()+$!%>
(+$)5+E!/'"#!45$*!"#(A67,2!5(!+5(+$!$+5(!b#'&52'25@+(!<+2$'672#()!*+$!V5)'(*!5(!*+$!<5(*+2'>
,67+!M&'2"5+$2!xcKRKOd[[d!k!dPPjJ!CDDIyE!/5+!$56725)+!-$gr+!#(*!-+,2'&2!*+,!V5)'(*+(!
,5(*J! +(2,M$+67+(*! *+,! O67&Q,,+&>O67&0,,>L$5("5M,! x:XOcU%PJ! GFH]yJ! *5+! 5(! *5+,+9! O67$522!
456725),2+(!W$52+$5+(E!!!
3'67!*+9!Docking!+$80&)2!+5(+!b#'(252'25@+!<+2$'672#()E!/5+,+!45$*!'&,!Scoring ;+"+567(+2!
xV%KcU et al.J! CDD`yE!U5+$;+5!45$*! *5+! 8$+5+!<5(*#(),+(27'&M5+! -J! 8Q$! *5+!<5(*#()! "45>
,67+(!V5)'(*!#(*!L$02+5(J!952!U5&8+!9'27+9'25,67+$!B0*+&&+!';)+,67A2"2E!/5+!K;,67A2"#()!
@0(! -! ;+$#72! *';+5! '#8! *+$! <+4+$2#()! *+$! _+67,+&45$1#()+(! "45,67+(! 8#(1250(+&&+(!
-$#MM+(!*+,!V5)'(*+(!#(*!*+,!L$02+5(,! xO[KUVJ!CDDDyE! X(,;+,0(*+$+! 85(*+(!75+$;+5!_',>
,+$,2088;$Q61+(;5(*#()+(J! 50(5,67+! _+67,+&45$1#()+(J! B+2'&&109M&+\+J! W'250(> >
_+67,+&45$1#()+(J! > >_+67,+&45$1#()+(!#(*!7N*$0M70;+!_+67,+&45$1#()+(!<+$Q61,567>
25)#()E!/+$!7N*$0M70;+!%88+12! 5,2! 59!_+,+(2&567+(!+(2$0M5,67+$!3'2#$J!*E7E! +,!7'(*+&2! ,567!
(5672!#9!+5(+!*5$+12+!_+67,+&45$1#()!xWV%<%J!CDDHyE!R5+&9+7$!;+$#72!*+$!+(2$0M5,67+!<+5>
2$')!*'$'#8J!*',,!,567!8Q$!*',!V5)'(*>L$02+5(>_',,+$>ON,2+9!*5+!^'7&!*+$!:$+57+52,)$'*+!'#8>
)$#(*!*+$!V5)'(*>K,,0"5'250(!'(!*',!L$02+5(!A(*+$2! x<ÑUB et al.J!CDDCn!WX[cU%3 et al.J!
CDD]n!WV%<%J!CDDHyE!!
!
HDRDP! G2;+*'25*2':!2%0!d-%9#,-5.#*++2,-.%!?9!O#;,2'("+(!"#!5*+(2585"5+$+(J!*5+!,2$#12#$+&&!A7(&567!"#!;+$+52,!)+8#(*+(+(!'125@+(!X(75;5>
20$+(!,5(*J!p+*067!+@+(2#+&&!+5(+!;+,,+$+!5(75;520$5,67+!K125@52A2!'#84+5,+(J!*5+(+(!a7(&567>
1+52,>!#(*!O#;,2$#12#$,#67+(E!<+5!a7(&5671+52,,#67+(!45$*!+5(!O'2"!@0(!c7'$'12+$5,251'!+5(+,!
K#,)'(),90&+1Q&,J! 952! *+(! B0&+1Q&+(! 5(! +5(+$! /'2+(;'(1! @+$)&567+(! x_XVV%[[J! CDDYyE!
/'$'#,!$+,#&25+$+(!B0&+1Q&+J!*5+!*+9!K#,)'(),90&+1Q&!67+95,67!A7(&567!,5(*J!';+$!*#$67'#,!
+5(+$!'(*+$+(!O2$#12#$1&',,+!'()+7g$+(!1g((+(E!!!
/5+! O#67+! ('67! O2$#12#$8$')9+(2+(! *')+)+(! ;+"+567(+2! 9'(! '&,! O#;,2$#12#$,#67+(!
xi?X3[?O et al.J! CDDHyE!<+5! *5+,+$!K$2! *+$! O#67+! +$7A&2!9'(!B0&+1Q&+J!4+&67+! *',!K#,>
)'(),90&+1Q&!'&,!<+,2'(*2+5&!57$+$!O2$#12#$!+(27'&2+(J!p+*067!@+$,675+*+(+!O#;,252#+(2+(!;+>
,52"+(E!!
EINLEITUNG
C`!
HDRDS! Q#'*2.99.+!Screening!
R5$2#+&&+, Screening!x<ÑUB!k!OcU3%X/%PJ!CDDDn!BcX33%OJ!CDDIy!;+"+567(+2!'&,!d;+$>
;+)$588!'&&+!B+270*+(!*+$!609M#2+$)+,2Q2"+(!O#67+! 5(!)$0r+(!O#;,2'("*'2+(;'(1+(J!*5+! 59!
U067*#$67,'2">R+$8'7$+(! *#$67)+8Q7$2! 4+$*+(! x_KV[%PO et al.J! GHHFyE! X(! *5+,+$! K$;+52!
4#$*+!Docking! '&,!<+4+$2#(),9+270*+! 8Q$!*',!@5$2#+&&+ Screening! +5()+,+2"2E!/',!@5$2#+&&+
Screening! A7(+&2! *'952! +5(+9!High Throughput Screening! e&';0$2+67(5,67+!U067*#$67,'2">
P+57+(#(2+$,#67#()f!xWV%<%J!CDD`n!OcU3%X/%P!k!:%cU3%PJ!CDDZyE!^5+&!*+,!@5$2#+&&+(
Screenings!5,2!+,J!'#,!+5(+$!)$0r+(!O#;,2'("*'2+(;'(1!*5+p+(5)+(!O#;,2'("+(!7+$'#,"#85&2+$(J!
*5+!'&,!M02+(25+&&+!_5$1,2088+!'9!#(2+$,#672+(!^5+&M$02+5(!+5(+!;50&0)5,67+!:#(1250(!;+,52"+(!
x_KV[%PO et al.J! GHHFyE! X9!K(,67&#,,! 1g((+(! *'((! *5+! 5(2+$+,,'(2+(J! M02+(25+&&+(!_5$1>
,2088+! 5(! @+$,675+*+(+(! L$Q8>odM2595+$#(),$#(*+(! 59! V';0$! 4+52+$! 0M2595+$2! 4+$*+(E! /5+!
B+270*+! +$&'#;2! +,! '#67! ,N(27+25,67! ,674+$! "#)A()&567+! O#;,2'("+(! 59! U067*#$67,'2">
@+$8'7$+(!'#8!57$+!_5$1#()!'9!^5+&M$02+5(!"#!#(2+$,#67+(!xBcX33%OJ!CDDIn!_KV[%PO et
al.J! GHHFyE! ^#,A2"&567! +$g88(+2! ,5+! *5+!Bg)&5671+52! +5(+,! +88+125@+(! [+,2,! 109;5('20$5,67+$!
O#;,2'(";5;&5027+1+(E!/5+!K("'7&!*+$!"#!,N(27+25,5+$+(*+(!O#;,2'("+(!1'((!*#$67!*',!@5$2#>
+&&+!Screening ,2'$1!)+,+(12!4+$*+(E!/'*#$67!4+$*+(!*5+!W0,2+(!5(!+5(+9!L$0p+12!)+,+(12E!!
/'!Docking! +5(+(! 707+(! P+67+('#84'(*! ;+*5()2J! ,5(*! @5$2#+&&+ Screening! O2#*5+(! )$0r+$!
O#;,2'("*'2+(;'(1+(! '#8! <',5,! @0(!Docking! ,+7$! "+52'#84+(*5)! x/UKVX_KV! k! cU%3J!
CDDHn!WPd%B%PJ! CDDIyE! L$0!O2$#12#$! 5,2!952! +5(+9!^+52'#84'(*!@0(! +5(5)+(!B5(#2+(! "#!
$+67(+(J!,0!*',,!,567!*+$!^+52'#84'(*!8Q$!+5(!@5$2#+&&+, Screening!+5(+$!)$0r+(!O#;,2'("*'>
2+(;'(1!&+5672!"#!+5(+$!;+2$A672&567+(!-$gr+!,#995+$2E!^#*+9!#98',,+(!@5$2#+&&+!O#;,2'(">
*'2+(;'(1+(!+5(+!*+#2&567!)$gr+$+!K("'7&!'(!O2$#12#$+(J!'&,!*5+!"#)+7g$5)+!$+'&+!O#;,2'(">
;'(1J!*'!'&&+!B0*5851'250(+(!*5+!5(!*+$!P+'&52A2!'#82$+2+(J!'&,!+5)+(,2A(*5)+!O2$#12#$+(!5(!*+$!
@5$2#+&&+(! O#;,2'("*'2+(;'(1! "#! +$8',,+(! ,5(*E! ^#$! +88+125@+(! <+'$;+52#()! *+,! @5$2#+&&+(!
Screenings! 5,2! *'7+$!9+5,2! *+$!%5(,'2"! +5(+$! &+5,2#(),8A75)+(J! M'$'&&+&+(!c09M#2+$M&'2280$9!
(024+(*5)!x/UKVX_KV!k!cU%3J!CDDHyE!^#$!%@'+$#()!*+,!@5$2#+&&+(!ScreeningsJ!,045+!
K#,4'7&! )++5)(+2+$!Docking>L'$'9+2+$J!45$*! "#(A67,2! @+$,#672J! *5+!d$5+(25+$#()! +5(+,!V5>
)'(*+(!*#$67!%5(M',,+(!5(!*+(!@09!V5)'(*+(!;+8$+52+(!L$02+5(1$5,2'&&!"#!$+M$0*#"5+$+(E!<+5!
*5+,+9! '&,! Re-Docking! ;+"+567(+2+(! R0$)+7+(! 1g((+(! 0M259'&+! %5(,2+&&#()! *+$!Docking!
L'$'9+2+$! 8Q$! *',! '(,67&5+r+(*+! @5$2#+&&+! Screening! +$9522+&2J! ,045+! *5+! ;+,2+! Scoring>
:#(1250(!8Q$!*5+,+!Docking>O2#*5+!;+,25992!4+$*+(E!/'$Q;+$!75('#,!+$&'#;2!*',!Re-DockingJ!
*+,!;+1'((2+(!V5)'(*+(!5(!*5+!U090&0)5+90*+&&+J!*5+!4+52+$+!R'&5*5+$#()!*+$!B0*+&&+E!/5+!
! ! EINLEITUNG
CI!
-Q2+!*+$!U090&0)5+90*+&&+!45$*!*';+5!Q;+$!*5+!<+$+67(#()!@0(!PBO/>_+$2+(!xcdU%3!k!
O[%P3<%P-J!GHFDy!*+$!V5)'(*'209&')+(!;+#$2+5&2E*
*
HDRDT! V/B:G#(29)*#$%.%!12'!".'.,-%2%3!0.'!&'.#.%!"#%02%3+.%.'3#.!/5+!Linear Interaction!Energy!B+270*+!eVX%f!*5+(2!*+$!<+$+67(#()!@0(!<5(*#(),'885(52A2+(!
xKiRXO[!k!BKP%VX?OJ!CDDGyE!/5+,+!,+95+9M5$5,67+!O59#&'250(,9+270*+!;',5+$2!'#8!B0>
&+1Q&*N('951>!0*+$!B0(2+>c'$&0>!)+(+$5+$2+(!%(,+9;&+,!@0(!W0(85)#$'250(+(!x:dVdLL%!k!
U?<<KP/J!CDD`yE!X(!*5+,+$!K$;+52!4#$*+!'#,,67&5+r&567!*5+!B0&+1Q&*N('951,59#&'250(!"#$!
-+(+$5+$#()!@0(!%(,+9;&+,!@+$4+(*+2E!/5+!B0&+1Q&*N('951!5,2!+5(+!,+95+9M5$5,67+!c09>
M#2+$,59#&'250(! *+$!<+4+)#()! @0(!K209+(! #(*!B0&+1Q&+(! '#8! *+$!<',5,! @0(!W$'828+&*+$(!
xWV%<%J! CDDHyE! <+5! *+$! %$9522&#()! *+$! K885(52A2+(! ('67! *+$! VX%>B+270*+! 45$*! @+$,#672J!
,5((@0&&+!X(80$9'250(+(!Q;+$!*5+!<5(*#(),+(+$)5+!'#,!O59#&'250(+(!*+$!M7N,51'&5,67!$+&+@'(>
2+(!^#,2A(*+!e8$+5!#(*!)+;#(*+(f!*+,!V5)'(*+(!';"#&+52+(!xKiRXO[ et al.J!CDDCyE!/5+!X*++!
75+$;+5! 5,2J!*',,!*5+! 8$+5+!<5(*#(),+(+$)5+!*+,!V5)'(*+(!'&,!a(*+$#()!,+5(+$! 8$+5+(!%(+$)5+!
;+2$'672+2!45$*J!4+((!+$!'#,!*+$!4A,,$5)+(!Vg,#()!e8$+5+$!^#,2'(*f!5(!,+5(+!,0&@'25,5+$2+!<5(>
*#(),2',67+! e)+;#(*+(+$!^#,2'(*f!Q;+$8Q7$2!45$*! x/%!KBdPXB et al.J!CDDFyE!K#,!*5+,+9!
-$#(*! ;+2$'672+2! *5+! VX%>B+270*+! (#$! *5+! ;+5*+(! %(*M#(12+! 59!V5)'(*+(;5(*#(),M$0"+,,!
e8$+5+$!^#,2'(*!,045+!)+;#(*+(+$!^#,2'(*fE!:Q$!*5+!90&+1Q&9+67'(5,67+(!<+$+67(#()+(!*+$!
<5(*#(),M$0"+,,+(*M#(12+!9Q,,+(!"4+5!VX%>O59#&'250(+(!*#$67)+8Q7$2!4+$*+(E!%5(+!O59#>
&'250(!8Q$!*+(!)+;#(*+(+(!V5)'(*+(!,045+!+5(+!4+52+$+!8Q$!*+(!V5)'(*+(!5(!4A,,$5)+$!Vg,#()!
e,5+7+!K;;5&*#()!GEGZfE!K#,!*+(!90&+1Q&9+67'(5,67+(!<+$+67(#()+(!+$7A&2!9'(!p+4+5&,!8Q$!
*5+!;+5*+(!^#,2A(*+!*5+!+&+12$0,2'25,67+(!,045+!*5+!@'(!*+$!_''&,!_+67,+&45$1#(),;+52$A)+E!!!
^#$!-+(+$5+$#()!+5(+,!VX%>B0*+&&,!4+$*+(!'#r+$*+9!+\M+$59+(2+&&+!<5(*#(),'885(52A2+(!952!
*+(!%(+$)5+2+$9+(!*+$!B0&+1Q&9+67'(51!109;5(5+$2J!#9!+5(+!<+4+$2#(),8#(1250(!"#$!R0$>
7+$,')+!@0(!<5(*#(),+(+$)5+(!"#!+$7'&2+(E!%,!&A,,2!,567!,0952!80&)+(*+!'&&)+9+5(+!3A7+$#()!
8Q$!*5+!<+,2599#()!*+$!8$+5+(!<5(*#(),+(+$)5+!'#8,2+&&+(!e,5+7+!-&+567#()!GEGf!xKiRXO[ et
al.J!CDDCyz!* !
! elsl
VDWsl UUG ! GEG!
!
U5+$;+5!5,2!S VDWslU T!)&+567!*+$!/588+$+("!*+$!%(,+9;&+9522+&4+$2+!*+$!R/_!_+67,+&45$>
1#()+(!59!)+;#(*+(+(!e&f!#(*!#()+;#(*+(+(!^#,2'(*!e,f!*+,!V5)'(*+(E!%(2,M$+67+(*!$+M$A>
,+(25+$2!S elslU Ö!*5+!+&+12$0,2'25,67+(!_+67,+&45$1#()+(!"45,67+(!V5)'(*!#(*!?9)+;#()E!
EINLEITUNG
CF!
/5+!-+45672#(),8'120$+(! J! !#(*! !4+$*+(!59!K(,67&#,,!*#$67!&5(+'$+!P+)$+,,50(!'(!+\M+>
$59+(2+&&!;+,25992+(!<5(*#(),+(+$)5+(!'()+M',,2E!!
!
Abbildung 1.15: Linear Interaction Energy Methode
/'$)+,2+&&2! ,5(*!*5+!(024+(*5)+(!O59#&'250(+(! 8Q$!*5+!<+,2599#()!*+$! 8$+5+(!<5(*#(),+(+$)5+!*+,!V5)'(*+(z!
"#9!+5(+(!*+$!V5)'(*!5(!Vg,#()!#(*!"#9!'(*+$+(!*+$!V5)'(*!)+;#(*+(!'(!*+(!@0&&!,0&@'25,5+$2+(!P+"+M20$E!!
:Q$!*5+!i#'&52A2!*+$!%$)+;(5,,+!;+5!*+$!O59#&'250(!952!Bd%!x722Mzoo444E67+9609ME609oyJ!
,M5+&2!*',!Setup!#(*!*5+!-Q2+!*+$!W$'828+&*M'$'9+2+$!8Q$!V5)'(*J!O0&@+(2!#(*!P+"+M20$!+5(+!
+(2,67+5*+(*+!P0&&+!xKVBVd: et al.J!CDD]yE!!
!
HDRDU! V.#*+*'25*2'$Y*#(#.'2%3!/5+!dM2595+$#()!@0(!V+52,2$#12#$+(! +$80&)2!9+5,2! 5(! +5(+9! &'()45+$5)+(J! 52+$'25@+(!L$0"+,,!
x3_KWK!k!PX/V%jJ!CDDYn!PKjBd3/ et al.J!CDDHyE!O5+!;+5(7'&2+2!(+;+(!*+$!R+$;+,,+>
$#()!*+$!<5(*#(),'885(52A2! "#!+5(+9!^5+&M$02+5(J!'#67!+5(+!dM2595+$#()!4+52+$+$!L'$'9+2+$!
45+!;+5,M5+&,4+5,+!Vg,&5671+52J
! ! EINLEITUNG
CH!
-$0;+!O2$#12#$>_5$1#(),;+"5+7#()+(! &',,+(!,567!'(7'(*!@0(!<5(*#(),'885(52A2+(!M02+(25+&>
&+$! X(75;520$+(! '#8,2+&&+(E! %,,+(25+&&+! _+67,+&45$1#()+(! "45,67+(! V5)'(*+(! #(*! L$02+5(!
1g((+(!*#$67!*',!%5(M',,+(!*+$!V5)'(*+(!5(!*5+!<5(*+2',67+!*+,!P+"+M20$,!eDockingf!+$952>
2+&2!4+$*+(E!O0!&',,+(!,567!;+5,M5+&,4+5,+!"#,A2"&567+!<+$+567+!5(!*+$!<5(*+2',67+!5*+(2585"5+>
$+(J!*5+!@0(!;+1'((2+(!V5)'(*+(!(067!(5672!'#,)+(#2"2!4+$*+(E!d*+$!+,!+$)+;+(!,567!,0!X*++(!
8Q$!*5+!R+$+5(8'67#()!@0(!V5)'(*+(J!#9!;+5,M5+&,4+5,+!*5+!Vg,&5671+52J!*5+!V5M0M75&5+!0*+$!
*5+! [$'(,M0$2>! #(*! R+$2+5&#(),+5)+(,67'82+(! eK/B%>%5)+(,67'82+(f! "#! 0M2595+$+(!
xOcU_KP/[ et al.J!CDDYyE!U5+$;+5!5,2!p+*067!*5+!W+((2(5,!*+,!2'2,A67&567+(!<5(*+90*#,!8Q$!
*5+! "#!0M2595+$+(*+!V+52,2$#12#$! +(2,67+5*+(*J!#9!@+$&A,,&567+!K#,,')+(!Q;+$!*5+!O2$#12#$>
_5$1#(),;+"5+7#()+(!2$+88+(!"#!1g((+(E!!!
%5(+! )$#(*&+)+(*+! P0&&+J! 5(! *+$! dM2595+$#()! @0(! X(75;520$+(! ^N1&5(>';7A()5+$! W5(',+(J!
,M5+&2!*5+!dM2595+$#()!75()+7+(*!*+$!O+&+125@52A2!*+$!X(75;520$+(!)+)+(Q;+$!'(*+$+(!W5(',+(J!
;"4E!)+)+(Q;+$!W5(',+(!*+,!_5$2,0$)'(5,9#,E!W5(',+(!,5(*!59!K&&)+9+5(+(!,+7$!7067!10(>
,+$@5+$2J! ,M+"5+&&! *5+! K[L><5(*+2',67+! 5,2! ;5,! '#8! 4+(5)+! K95(0,A#$+'#,2'#,67+! 5*+(25,67!
xWV%<%J!CDDHn!W?<X3jX!k!BsVV%PJ!CDD]n!3d<V% et al.J!CDD]yE!/5+,!9'672!+,!,6745+>
$5)!,+&+125@+!W5(',+>X(75;520$+(!"#!+(24561+&(E!=+*067!1'((!;+$+52,!+5(!+5("5)+$!K#,2'#,67!5(!
*+$! K[L><5(*+2',67+! )+(#2"2! 4+$*+(J! #9! ;+5,M5+&,4+5,+! +5(+! "#,A2"&567+! <5(*+$+)50(! "#!
'*$+,,5+$+(!e-&5@+6J!K<V>W5(',+f!xW?<X3jX!k!BsVV%PJ!CDD]yE!_+52+$75(!,M5+&2!082!'#67!
*5+!Vg,&5671+52!@0(!W5(',+>X(75;520$+(!+5(+!)$0r+!P0&&+! 59!V+52,2$#12#$0M2595+$#(),M$0"+,,!
x=K?[%VK[ et al.J!CDDZn!WUXP% et al.J!CDD]n!dOVd< et al.J!CDDHyE!!
HDe! W2&3);.%+*.992%3!/5+! $','(2+!_+52+$+(24561&#()! @0(!c09M#2+$7'$*4'$+! #(*!O0824'$+! +$&'#;2! +,! 7+#2+J! ,+7$!
)$0r+!/'2+(9+()+(!;50>!#(*!67+905(80$9'25,67!5((+$7'&;!1#$"+$!^+52!"#!@+$'$;+52+(E!c09>
M#2+$)+,2Q2"2+!B+270*+(!*+,!@5$2#+&&+(!Screenings!#(*!*+,!$'250('&+(!Drug Designs!4+$*+(!
9522&+$4+5&+! $0#25(+9Ar5)! 5(! *+$! K)$'$>J! [5+$)+,#(*7+52,>! #(*! 7#9'(+(! L7'$9'80$,67#()!
+5()+,+2"2E! %5(! 456725)+$! <+$+567! *+$! @+2+$5(A$9+*5"5(5,67+(! _5$1,208880$,67#()! ,5(*! *5+!
L'$',520,+(!@0(!U'#,>!#(*!3#2"25+$+(E Eimeria tenella!5,2!+5(!M'$',525,67!&+;+(*+(+$!L$020"0+!
*+,! O2'99+,! *+$! ApicomplexaE! /5+,+$! 25+$M'270)+(+! L'$',52! @+$#$,'672! W01"5*50,+J! +5(+!
UQ7(+$+$1$'(1#()!*5+!4+&24+52! "#!707+(!R+$&#,2+(! 5(!*+$!-+8&Q)+&M$0*#1250(! 8Q7$2E! X(!*+$!
@0$&5+)+(*+(! K$;+52! ,0&&2+! *5+! _5$1,208885(*#()! @0(! 67+9027+$'M+#25,67+(! K(25101"5*5'!
*#$67!90*+$(+!B+270*+(!*+$!<50>!#(*!c7+95+5(80$9'251!*#$67)+8Q7$2!4+$*+(E!X(!@0$'()+>
EINLEITUNG
YD!
)'()+(+(!%\M+$59+(2+(!4#$*+(!c/W>L$02+5(+!'&,!)++5)(+2+!^5+&M$02+5(+!5(!E. tenella!;+$+52,!
67+95,67! @'&5*5+$2! e,5+7+! [';+&&+! GEYfE! K#r+$*+9! 10((2+(! ;+$+52,! +$,2+! X(75;520$+(! *+$!
%2cPWC! l! +5(+$! c/W! '#,! E. tenella! l! 5(! +5(+9! @5$2#+&&+(! Screening! 5*+(2585"5+$2! 4+$*+(!
x<%j%PJ!CDDIyE!!!
/5+! @0$&5+)+(*+! K$;+52! )&5+*+$2! ,567! 5(! *5+! 80&)+(*+(! *$+5! K;,67(522+z! GE! <505(80$9'25,67+!
X*+(2585"5+$#()! #(*!c7'$'12+$5,5+$#()! @0(!c/W!L$02+5(+(! #(*! *+$+(!K125@'20$+(! e^N1&5(+f!
'#,!E. tenellaE!CE!K($+567+$#()!#(*!dM2595+$#()!@0(!%2cPWC>X(75;520$+(!,045+!YE!R5$2#+&&+,!
Screening!952!*+$!Docking>O0824'$+!LVK3[O!('67!L8BPW>!#(*!%2BPW>X(75;520$+(E!!
^5+&! 59! +$,2+(!K;,67(522! *5+,+$!K$;+52!4'$! +5(+! ;505(80$9'25,67+! [5+8+('('&N,+! @0(!c/W>
7090&0)+(! ,045+! ^N1&5(>A7(&567+(! L$02+5(+(! @0(! E. tenellaE! U5+$;+5! ,0&&2+! "#(A67,2! +5(+!
c7'$'12+$5,5+$#()!*5+,+$! 7090&0)+(!c/W,! 5(!<+"#)! '#8!:#(1250(!#(*!O2$#12#$! ,2'2285(*+(E!
/5+,+!c7'$'12+$5,5+$#()!+$80&)2+!'(7'(*!*+$!O'99&#()!#(*!<+4+$2#()!@0(!O+b#+("5(80$9'>
250(E!c/W,!,5(*!5((+$7'&;!*+$!Apicomplexa!8Q$!Plasmodium falciparumJ!*+,,+(!-+(09!@0&&>
,2A(*5)! ,+b#+("5+$2! 5,2J! '9! ;+,2+(! #(2+$,#672E! /'7+$! 4#$*+! *5+! O#67+! 952! *+(! ;+1'((2+(!
c/W,!#(*!^N1&5(+(!@0(!P. falciparum *#$67)+8Q7$2E!/'$Q;+$!75('#,!,0&&2+(!*5+!5*+(2585"5+$>
2+(!^N1&5(+!*+(!c/W!L$02+5(+(!'#,!E. tenella!'&,!M02+(25+&&+!K125@'20$+(!"#)+0$*(+2!4+$*+(E!!
/',!"4+52+!^5+&!*5+,+$!K$;+52!4'$!*5+!K($+567+$#()!@0(!X(75;520$+(J!*5+!;+$+52,!+5(+!K125@52A2!
'#8!%2cPWC!)+"+5)2!7';+(E!/5+,+!X(75;520$+(!,2'992+(!'#,!+5(+9!8$Q7+$+(!@5$2#+&&+(!Scree-
ning!x<%j%PJ!CDDIy!#(*!10((2+(!;5067+95,67!;+,2A25)2!4+$*+(E!X(,)+,'92!4'$+(!"#!<+)5((!
*5+,+$!K$;+52!+&8!O#;,2'("+(!;+1'((2J!*5+!+5(+(!XcZD>_+$2!@0(!1&+5(+$!GD!ÜB!"+5)2+(E!^#*+9!
4'$! +,!^5+&! *5+,+$!K$;+52J! *5+!<+("595*'"0&>c'$;0(52$5&+J! *5+! in vitro! '&,!V+52,2$#12#$+(!*+$!
%2cPWC! ;+,2A25)2! 4+$*+(! 10((2+(J in silico! "#! 0M2595+$+(E! U5+$8Q$! ,0&&2+! *+$! <5(*+90*#,!
*5+,+$!O2$#12#$1&',,+!59!/+2'5&!'('&N,5+$2!4+$*+(J!#9!59!K(,67&#,,!)++5)(+2+!B0*5851'250(+(!
*5+,+$!R+$;5(*#()+(!@0$,67&')+(!"#!1g((+(E!!!
X9!*$522+(!K;,67(522!*5+,+$!K$;+52!,0&&2+!+5(!@5$2#+&&+,!Screening!('67!M02+(25+&&+(!X(75;520$+(!
*+$! L8BPW! #(*! %2BPW! *#$67)+8Q7$2! 4+$*+(E! /5+,+! L$02+5(+! ,5(*! +;+(8'&&,! ^N1&5(>
';7A()5)+!W5(',+(!*+$!d$)'(5,9+(!P. falciparum!#(*!E. tenellaE!U5+$"#!,0&&2+!*5+!Docking>
O0824'$+! LVK3[O! eL$02+5(>V5)'(*>K3[>ON,2+9f! @+$4+(*+2! #(*! +@'+$2! 4+$*+(E! 3'67!
B+270*+(+@'+$#()!,0&&2+!"#*+9J!)++5)(+2+!O#;,2$#12#$+(!'#,!+5(+$!O2$#12#$*'2+(;'(1!'&,!
M02+(25+&&+!Hits!'#,)+4A7&2!4+$*+(J!#9!+5(+!V5,2+!)++5)(+2+$!X(75;520$+(!7#9'(+$!W5(',+(!"#!
+$7'&2+(J!*5+!5(!4+52+$+(!O67$522+(!8Q$!*5+!M'$',52A$+(!^5+&M$02+5(+!0M2595+$2!4+$*+(!1g((+(E!
! ! MATERIAL UND METHODEN
YG!
J!I)*.'#)9!2%0!I.*-$0.%!!
JDH! X)'0E)'.!2%0!G$&*E)'.!!
JDHDH! X)'0E)'.!!
* V5(#\>c&#,2+$z!! O#(!:5$+!RCD"J!
! ! ! ! C\!KB/!dM2+$0(!CZD!eYC!NodesfJ!
! ! ! ! <+2$5+;,,N,2+9z!O?O%!V5(#\!HED!
* Lcz! ! ! /+&&!dM25L&+\!-}GGDJ!\F`!
! ! ! ! <+2$5+;,,N,2+9z!B56$0,082!_5(*04,!}L!eOLGf! !!
* ?(5\>c09M#2+$z! O-X!d$5)5(!YCDDJ!O-X!dCJ!!
! ! ! ! O-X!d62'(+!CJ!O-X!:#+&! !
! ! ! ! <+2$5+;,,N,2+9z!X$5\!`EZ!
!
JDHDJ! G$&*E)'.!!
* <50%*52!ZEDEHJ O+b#+(">Alignment!%*520$
30$27!c'$0&5('!O2'2+!?(5@+$,52NJ!?OKJ!xUKVVJ!CDDHy!!
* c&#,2'&_!@GE]J!L$0)$'99!"#$!%$,2+&&#()!9#&25M&+$J!)&0;'&+$!O+b#+(">Alignments!
%#$0M+'(!B0&+6#&'$!<50&0)N!V';0$'20$NJ!U+5*+&;+$)J!x[UdBLOd3 et al.J!GHH]y!!
* c+$5#,C!@E]EGDJ!c7+905(80$9'25,67+,!O0824'$+M'1+2!
K66+&$N,!X(6EJ!O'(!/5+)0J!?OKJ!x722Mzoo444E'66+&$N,E609oy!!
* c0$5('!YEYJ!W0(80$9+$)+(+$'20$!
! B0&+6#&'$!3+240$1,!-9;UJ!%$&'()+(!x722Mzoo444E90&>(+2E609y!!
* -+(+/06!CE`EDDGJ!O+b#+(">Alignment!%*520$!
L522,;#$)7!O#M+$609M#25()!c+(2+$J!?OKJ!x3XcUdVKO et al.J!GHHIy!!
* -dV/!CEC!;5,!]EDJ!O0824'$+!"#9!L$02+5(>V5)'(*>Docking
! c'9;$5*)+!c$N,2'&&0)$'M756!/'2'!c+(2$+J!?WJ!x=d3%O et al.J!GHHZy !
MATERIAL UND METHODEN
YC!
* X(,5)72!XXJ!c7+905(80$9'25,67+,!O0824'$+M'1+2!
K66+&$N,!X(6EJ!O'(!/5+)0J!?OKJ!x722Mzoo444E'66+&$N,E609oy!!
* XOXOo<',+J!/'2+(;'(1,N,2+9!8Q$!67+95,67+!O#;,2'("+(!
! B/V!X(80$9'250(!ON,2+9,!X(6EJ!O'(!P'90(J!?OKJ!x722Mzoo444E9*&E609y!!
* B+2'&58+![$5(52NGE]J!<505(80$9'25,67+,!L$0)$'99M'1+2!
B+2'&58+!K-J!_5(*+(J!x722Mzoo444E9+2'&58+E*+oy!!
* Bd%J!c7+905(80$9'25,67+,!O0824'$+M'1+2!
[$5M0,!X(6EJ!O2E!V0#5,J!?OKJ!x722Mzoo444E67+9609ME609oy!!
* B0&6'*J!O0824'$+!"#$!*$+5*59+(,50('&+(!R5,#'&5,5+$#()!@0(!B0&+1Q&+(!
[$5M0,!X(6EJ!O2E!V0#5,J!?OKJ!x722Mzoo444E2$5M0,E609y!
x<PXcWBK33J!GHHZn!<PXcWBK33 et al.J!CDDDy!!
* L5M+&5(+L5&02J!c7+905(80$9'25,67+,!L5M+&5(+200&!
K66+&$N,!X(6EJ!O'(!/5+)0J!?OKJ!x722Mzoo444E'66+&$N,E609oy!!
* R0&,#$8tJ!O0824'$+M'1+2!"#$!<+$+67(#()!@0(!/+,1$5M20$+(!
! B0&+6#&'$!/5,60@+$N!V2*EJ!X[KVjJ!x722Mzoo444E90&*5,60@+$NE609y!
! ! ! !
JDJ! A)*.%;)%5.%!
Tabelle 2.1: Verwendete Datenbanken. !
A)*.%;)%5.%! LFV! F.&.'.%1!
<5(*5()/<! 722Mzoo444E;5(*5()*;E0$)! xcU%3 et al.J!CDDGy!
/?/! 722Mzoo*#*E*0615()E0$)o! xU?K3- et al.J!CDD`y!
-+(<'(1! 722Mzoo444E(6;5E(&9E(57E)0@o! x<%3Od3 et al.J!GHHYy!
-+(O6'(! 722Mzoo)+(+,E952E+*#o-%3OcK3E729&x<?P-%!k!WKPVX3J!
GHHIy!
L#;B+*! 722Mzoo444EM#;9+*E)0@o! x_U%%V%P et al.J!CDD]y!
! ! MATERIAL UND METHODEN
YY!
A)*.%;)%5.%! LFV! F.&.'.%1!
LPdOX[%! 722Mzoo#,E+\M',NE0$)oM$0,52+o! xU?Vd et al.J!CDD`y!
L8'9!722Mzoo444E,'()+$E'6E#1oO0824'$+o!
L8'9o5(*+\E,729&!x<K[%BK3 et al.J!CDD]y!
X(2+$L$0! 722Mzoo444E+;5E'6E#1o5(2+$M$0o! xB?V/%P et al.J!CDDZy!
O45,,>L$02! 722Mzoo#,E+\M',NE0$)o,M$02!x<d%cWBK33 et al.J!
CDDYy!
L/<! 722Mzoo444E$6,;E0$)oM*;o! x<%PBK3 et al.J!CDDYy!
!
JDP! I.*-$0.%!12(!G.f2.%16.'39.#,-!JDPDH! G.f2.%1:Alignments!
O+b#+(">Alignments!*5+(+(!*+$!/+2+1250(!#(*!/'$,2+&&#()!10(,+$@5+$2+$! ,045+!*5@+$,+$!<+>
$+567+! "45,67+(! "4+5! eM''$4+5,+,! Alignmentf! 0*+$! 9+7$+$+(! O+b#+("+(! e9#&25M&+, A-
lignmentfE!/5+!@0$&5+)+(*+(!9#&25M&+(!AlignmentsJ!"#$!%$,2+&&#()!*+$!;505(80$9'25,67+(![5+>
8+('('&N,+J! 4#$*+(! '#,,67&5+r&567! 952! *+9! 5(! *+$! O0824'$+! Metalife! Trinity 1.4! 59M&+>
9+(25+$2+(!L$0)$'99!ClustalW!x[UdBLOd3 et al.J!GHH]yJ!#(2+$!R+$4+(*#()!*+$!O2'(*'$*>
+5(,2+&&#()+(J!*#$67)+8Q7$2E!U5+$"#!4#$*+(!*5+!O+b#+("+(!5(!+5(!O+b#+(",+2!59!B0*#&!Meta-
life-Predictor!*+$!O0824'$+!Metalife!Trinity 1.4 xB+2'&58+!K-J!_5(*+(J!/+#2,67&'(*y 59M0$>
25+$2E!ClustalW +$,2+&&2! '#,!9#&25M&+(!:KO[K>/'2+5+(!)&0;'&+J!9#&25M&+!AlignmentsE!/5+!%$>
)+;(5,,+!4#$*+(J!"#$!R5,#'&5,5+$#()!#(*!)$'M75,67+(!P+M$0*#1250(J!5(!*',!L$0)$'99!Gene-
Doc!x3XcUdVKO et al.J!GHHIy!59M0$25+$2E!
!
JDS! /0.%*##.'2%3!6$%!4A=:<-%9#,-.%!2%0!7859#%:<-%9#,-.%!>'$*.#%.%!K&&+! O+b#+("*'2+(! 4#$*+(! @09! 3'250('&! c+(2+$! 80$! <502+67(0&0)N! X(80$9'250(! e3c<XJ!
722Mzoo444E(6;5E(&9E(57E)0@of!0*+$!@09!O'()+$!X(,252#2+!e722Mzoo444E,'()+$E'6E#1of!;+"0)+(E!
^#$!X*+(2585"5+$#()!@0(!c/W>A7(&567+(!L$02+5(+(J!,045+!*+$!+(2,M$+67+(*+(!^N1&5(+!'#, E.
tenellaJ 4#$*+(! $+1#$,5@+!<VKO[>,#67+(! e<VKO[3J
MATERIAL UND METHODEN
Y]!
E. tenella!-+(09,E!P+1#$,5@+!<VKO[>O#67+(!4#$*+(!952!;+$+52,!;+1'((2+(!c/W>A7(&567+(!
#(*! ^N1&5(>A7(&567+(! L$02+5(+(! @0(!P. falciparum! *#$67)+8Q7$2E! K#r+$*+9! 4#$*+(! "#@0$!
5*+(2585"5+$2+!O+b#+("+(!@0(!'(*+$+(!d$)'(5,9+(!'#,!*+9!O2'99!*+$!ApicomplexaJ!45+!;+5>
,M5+&,4+5,+! Plasmodium yoelii yoeliiJ!Cryptosporidium parvumJ!Cryptosporidium hominisJ!
Theileria parvaJ!Theileria annulata! #(*!Toxoplasma gondii! x%3-%VOJ!CDDZyJ! "#$!4+52+$+(!
R+$585"5+$#()!*+$!9#29'r&567+(!-+(,2$#12#$!9522+&,![<VKO[3!7+$'()+"0)+(E!/+$!R+$)&+567!
*+$! ;+1'((2+(! L$02+5(+! +$&'#;2+! (+;+(! +5(+$! )+('#+(!c7'$'12+$5,5+$#()J!45+! ;+5,M5+&,4+5,+!
*+$! %$9522&#()! @0(! O+b#+("9025@+(! #(*! ,2$#12#$+&&+(! %5)+(,67'82+(J! '#67! *5+! <+$+67(#()!
*+,!W0(,+$@5+$#(),)$'*+,!*5+,+$!L$02+5(+J!5((+$7'&;!*+$!+5("+&(+(!d$*(#()+(!CoccidiaJ!Hae-
mosporida!#(*!PiroplasmidaE!/5+!5*+(2585"5+$2+(!O+b#+("+(!'#,!E. tenellaJ!952!+5(+9!E-value!
xWdP:J!CDDYy!1&+5(+$!GD>Y!#(*!+5(+9!Bit-Score!)$gr+$!'&,!GDDJ!4#$*+(!'&,!,5)(5851'(2!+$'67>
2+2E!!!
/'!+,!,567!;+5!*+(!,0!+$9522+&2+(!(+#+(!O+b#+("+(!#9!)+(095,67+!O+b#+("+(!7'(*+&2J!9#,,>
2+(! 59! 3'67)'()! (067! %\0(>! #(*! X(2$0(;+$+567+! @0$7+$)+,')2! 4+$*+(E! /'"#! 4#$*+! *+$!
GenScan>O+$@+$!*+,!Massachusetts Institute of Technology!@+$4+(*+2E!/',!L$0)$'99!GenS-
can! x<?P-%!k!WKPVX3J!GHHIy!;',5+$2!'#8!+5(+9!U5**+(!B'$10@!B0*+&&! x%//jJ!CDD]yJ!
*',! 2$'5(5+$2!4#$*+J!*5+!%\0(>oX(2$0(,2$#12#$!+#1'$N025,67+$!-+(+!'#,!)+(095,67+(!O+b#+(>
"+(!"#!+\2$'75+$+(E!/5+,+!R0$7+$,')+!@+$&5+8!(5672!8+7&+$8$+5J!,0!*',,!*5+!%\0(>oX(2$0()$+("+(!
9'(#+&&J!9522+&,!*+$!-[>K->P+)+&! xBd?3[J!GHFCy!'()+M',,2!4+$*+(!9#,,2+(E!%5(!9g)&5>
67+$!-$#(*! 8Q$!*5+! 8+7&+$7'82+!R0$7+$,')+J! 5,2! *5+!VA()+!*+$! +5()+,+2"2+(!)+(095,67+(!O+>
b#+("+(J!*5+! 2+5&4+5,+!9+7$+$+! 2'#,+(*!<',+(M''$+!#98',,2+E!%5(+!10$$+12+!R0$7+$,')+!*+$!
%\0(oX(2$0(>O2$#12#$!45$*J!952!"#(+79+(*+$!VA()+!#(*!W09M&+\52A2!*+$
! ! MATERIAL UND METHODEN
YZ!
,#()!@0(!CJG!hJ!4#$*+!*5+,+!O2$#12#$!'&,![+9M&'2!8Q$!*5+!B0*+&&;5&*#()!+5()+,+2"2E!/5+!K#,>
4'7&!*+$![+9M&'2,2$#12#$!+$80&)2+!'(7'(*!@+$,675+*+(+$!L'$'9+2+$J!45+!;+5,M5+&,4+5,+!5,02$0>
M+$! /+;N+>_'&&+$>[+9M+$'2#$8'120$+(! e<>:'120$+(f! x-Kd! k! L%3-J! GHHHy! *+$! [+9M&'2>
O2$#12#$+(J!K#,$5672#()!*+$!O+52+(1+22+(J!,045+!<+$+67(#()!*+$!9522&+$+(!b#'*$'25,67+(!K;>
4+567#()!*+$!+5("+&(+(!K209&')+(!eRoot Mean Square DeviationJ!PBO/>_+$2+f!xcdU%3!
k! O[%P3<%P-J! GHFDy! "45,67+(! Template>! #(*! Target>O2$#12#$E! %,! 4#$*+(! p+4+5&,! 8Q$!
%2BPW! #(*! L8BPW! "+7(!U090&0)5+90*+&&+!952! *+9! 7g67,29g)&567+(!dM2595+$#(),)$'*!
)+(+$5+$2E!=+*+,!B0*+&&!4#$*+!"#(A67,2!)$0;!95(595+$2J!"#$!dM2595+$#()!*+$!,2+$5,67+(!X(2+$>
'1250(+(J!#(*!*'((! p+4+5&,!*',!;+,2+!^45,67+(M$0*#12!(067!+5(9'&!+24',! 8+5(+$!95(595+$2E!
^#$!B5(595+$#()!4#$*+!*',!BB::H]!W$'828+&*!xUKV-P%3J!GHH`y!@+$4+(*+2E!%5(5)+!O+5>
2+(1+22+(!4#$*+(!)+)+;+(+(8'&&,!9'(#+&&!0M2595+$2E!U5+$8Q$!4#$*+!;+5,M5+&,4+5,+!+5(!)++5)>
(+2+$+,!P02'9+$!;+(#2"2E!/5+,!*5+(2+!*+$!dM2595+$#()!*+$!<5(*#(),90*5!+5("+&(+$!P+8+$+(">
&5)'(*+(E! /5+! R5,#'&5,5+$#()! *+$! 85('&+(! B0*+&&+! +$80&)2+! 952! *+9! L$0)$'99! B0&6'*!
x<PXcWBK33J!GHHZn!<PXcWBK33 et al.J!CDDDyE!:Q$!*5+!-+(+$5+$#()!*+$!P'9'67'(*$'(>
L&02,!xPKBKcUK3/PK3 et al.J!GH`Yy!4#$*+!+;+(8'&&,!*',!O0824'$+M'1+2!Bd%!@+$4+(*+2E!!
!
JDU! G2;+*'25*2':!2%0!d-%9#,-5.#*++2,-.%!!?9!O#;,2'("+(!"#!5*+(2585"5+$+(J!*5+!,2$#12#$+&&!A7(&567!"#!;+$+52,!;+1'((2+(!'125@+(!X(75;520>
$+(!,5(*!x<%j%PJ!CDDIyJ!';+$!+@+(2#+&&!+5(+!;+,,+$+!5(75;520$5,67+!K125@52A2!'#84+5,+(J!4#$>
*+(!a7(&5671+52,>!#(*!O#;,2$#12#$,#67+(!*#$67)+8Q7$2E!U5+$8Q$!4#$*+!"#9!+5(+(!*',!67+95>
,67+!/'2+(;'(1,N,2+9!XOXOo<',+!@+$4+(*+2!#(*!"#9!'(*+$+(!*5+!L5M+&5(+L5&02!O0824'$+E!<+5!
a7(&5671+52,,#67+(!45$*!+5(!O'2"!@0(!c7'$'12+$5,251'!+5(+,!K#,)'(),90&+1Q&,!952!*+(!B0>
&+1Q&+(!5(!+5(+$!/'2+(;'(1!@+$)&567+(E!^#$!<+,67$+5;#()!*+$!67+95,67+(!O2$#12#$+(!4#$*+(!
*5+! 5(!L5M+&5(+L5&02! 59M&+9+(25+$2+(!Fingerprints! xK66+&$N,!X(6EJ!O'(!/5+)0J!?OKy!%c:Lá]!
#(*!:c:Lá]!xPd-%PO et al.J!CDDZy!;+$+67(+2E!%,!4#$*+!+5(!['(59020>X(*+\!x=KccKP/J!
GHGCn!_XVV%[[J!CDD`y!@0(!IDq!'&,!O674+&&+(4+$2! 8Q$!*5+!a7(&5671+52,,#67+(!@+$4+(*+2E!
%,!$+,#&25+$2+(!75+$'#,!CEYYG!O#;,2'("+(J!*5+!*+9!K#,)'(),90&+1Q&!67+95,67!A7(+&2+(J!2+5&>
4+5,+!';+$!'#67!+5(+$!'(*+$+(!O2$#12#$1&',,+!'()+7g$2+(E!!!/5+!O#67+!('67!O2$#12#$8$')9+(2+(!+$)';!]CEDHH!O#;,2'("+(E!<+5!*5+,+$!K$2!*+$!O#67+!+$7A&2!
9'(!B0&+1Q&+J! 4+&67+! *',! K#,)'(),90&+1Q&! '&,! <+,2'(*2+5&! 57$+$! O2$#12#$! +(27'&2+(J! ';+$!
@+$,675+*+(+!O#;,252#+(2+(!2$')+(E!!
!
MATERIAL UND METHODEN
Y`!
JDc! Q$';.'.#*2%3!0.'!G2;+*)%10)*.%!&N'!0)+!($9.529)'.!A$,5#%3!!c7+95,67+! R+$;5(*#()+(! 1g((+(! 5(! *+$! P+'&52A2! @+$,675+*+(+! ^#,2A(*+J! 75(,5672&567! 57$+$!
,2$#12#$+&&+(! %5)+(,67'82+(! eW0(80$9+$+fJ! 57$+$! +&+12$067+95,67+(! %5)+(,67'82+(! eX0(5,5+>
$#(),"#,2A(*+f!#(*!57$+,!['#209+$5,5+$#(),"#,2'(*+,!e['#209+$+f!xd%VVX%3 et al.J!CDD`yJ!
'((+79+(E!/5+!@+$,675+*+(+(!W0(80$9+$+!+5(+$!67+95,67+(!R+$;5(*#()!1g((+(!59!90&+1#>
&'$+(!Docking!'#209'25,67!;+2$'672+2!4+$*+(E!Bg)&567+!X0(5,5+$#(),"#,2A(*+!#(*!['#209+$+!
9Q,,+(! p+*067! @0$7+$! ;+$+67(+2! #(*! '&,! 5(*5@5*#+&&+! O2$#12#$+(! ';)+,M+567+$2!4+$*+(E!/5+!
K#8;+$+52#()!*+$!O#;,2'("*'2+(;'(1!4#$*+!952![6&>O1$5M2+(J!#(2+$!%5(,'2"!*+$!L$0)$'995+$>
;5;&5027+1!c'62@,!xXUV%3:%V/[ et al.J!CDDCn!d%VVX%3 et al.J!CDDZyJ!*#$67)+8Q7$2E!:Q$!*5+!
,0!@0$;+$+52+2+(!O2$#12#$+(!4#$*+(!*'((!59!K(,67&#,,!O2'$210(80$9+$+!952!*+$!O0824'$+!c0>
$5('! !;+$+67(+2E!^#*+9!4#$*+(!*5+!O2$#12#$+(!+(2,M$+67+(*!57$+,!X0(5,5+$#(),"#,2'(*+,!952!
_',,+$,2088'209+(!';)+,A225)2E!
!
JDR! Docking JDRDH! KZVA!K&,! L$0)$'99! "#9! /#$678Q7$+(! *+,! DockingsJ! "#$! <+4+$2#()! *+$! M02+(25+&&+(! %2cPWC!
xWX33KXP/ et al.J!CDD]y!X(75;520$+(J!4#$*+!-dV/!@]ED!eGenetic Optimisation for Ligand
Dockingf! x=d3%O et al.J! GHHZy! @+$4+(*+2E! -dV/! ;+(#2"2! +5(+(! )+(+25,67+(! K&)0$5279#,!
"#9!%5(M',,+(!*+$!67+95,67+(!O2$#12#$+(!5(!*5+!L$02+5(;5(*+2',67+E!<+5!+5(+9!)+(+25,67+(!
K&)0$5279#,!45$*!*+$!L$0"+,,!*+$!%@0ú(!9522+&,!B#2'250(J!P+109;5('250(!#(*!O+&+1250(!
@0(!X(*5@5*#+(!+5(+$!L0M#&'250(!,59#&5+$2E!/5+!X(*5@5*#+(!+(2,M$+67+(!75+$;+5!*+(!p+4+5&5)+(!
V5)'(*+(10(80$9'250(+(E!%5(+!L0M#&'250(!5,2!*+9('67!*5+!B+()+!'&&+$!V5)'(*+(10(85)#$'25>
0(+(E!/5+!:52(+,,!eScoring>_+$2f!+5(+,!X(*5@5*##9,!;"4E!+5(+$!V5)'(*+(10(80$9'250(J!45$*!
9522+&,!dM2595+$#(),1$52+$5+(J!45+!"E!<E!*+$!_+67,+&45$1#(),+(+$)5+!"45,67+(!+5(+9!V5)'(*!
#(*!,+5(+9!L$02+5(J!;+$+67(+2E!/',!L$02+5(!45$*!;+5!*5+,+9!R+$8'7$+(!,2'$$!)+7'&2+(J!(#$!*5+!
L0,5250(+(!*+$!_',,+$,2088'209+!1g((+(!,567!A(*+$(E!/+$!V5)'(*!75()+)+(!45$*!8&+\5;+&!;+>
7'(*+&2E!:Q$!*',!Docking!1g((+(!5(!-dV/!@+$,675+*+(+!K$2+(!@0(!Scoring>:#(1250(+(!'#,>
)+4A7&2! 4+$*+(E! 3+;+(! *+(! ;+$+52,! 59M&+9+(25+$2+(! Scoring>:#(1250(+(! -dV/>Score!
x=d3%O et al.J!GHHZy!#(*!c7+9>Score!x%V/PX/-% et al.J!GHHIy!5,2!+,!9g)&567J!;+(#2"+$*+>
85(5+$2+!:#(1250(+(!"#!(#2"+(E!:Q$!*',!,0!*#$67)+8Q7$2+!Docking!4#$*+!*5+!-dV/>Score!:52>
(+,,8#(1250(!@+$4+(*+2E!/5+,+!+(27A&2!80&)+(*+!8Q(8!<+4+$2#(),109M0(+(2+(z!GE!<5(*#(),>
+(+$)5+!*+$!_',,+$,2088;$Q61+(!"45,67+(!L$02+5(!#(*!V5)'(*! e5(2+$90&+1#&'$+!_',,+$,2088>
;$Q61+(fJ!CE! 5(2+$90&+1#&'$+!@'(>*+$>_''&,>%(+$)5+! e@*4f!"45,67+(!L$02+5(!#(*!V5)'(*J!YE!
! ! MATERIAL UND METHODEN
YI!
5(2$'90&+1#&'$+! @*4>%(+$)5+! *+,!V5)'(*+(J!]E!%(+$)5+!*+$
MATERIAL UND METHODEN
YF!
B#%+*.992%3.%! >)')(.*.';.1.#,-%2%3!#%!KZVA! @.'*!
M'$'9á85&+! *+8'#&2!
,+2á&5)'(*á'209á85&+,! G!
,+2áM$02+5(á'209á85&+,! G!
/'2'!:5&+,!
20$*5,2á85&+! *+8'#&2!
*5,M&'N! D!
5(2+$('&á&5)'(*á7á;0(*,! G!
(á&5)'(*á;#9M,! D!
8&5Má8$++á60$(+$,! G!
8&5Má'95*+á;0(*,! G!
8&5MáM&'('$á(! G!
8&5MáMN$'95*'&á(! G!
$02'2+á6'$;0\N&56á07! 8&5M!
:&'),!
#,+á20$*5,2! G!
[+$95('250(! +'$&Ná2+$95('250(! D!
c0(,2$'5(2,! 80$6+á60(,2$'5(2,! D!
5(525'&á@5$2#'&áM2á9'267á9'\! YED!
$+&'25@+á&5)'(*á+(+$)N! D!
:52(+,,!:#(6250(!
,2'$2á@*4á&5(+'$á6#2088! `!
!
JDRDJ! >VW?MG!/',!Docking>L$0)$'99!LVK3[O!eProtein Ligand Ant Systemf!xWdP< et al.J!CDD`n!WdP<
et al.J!CDDHy!*5+(2+!"#9!@5$2#+&&+(!Screen!('67!M02+(25+&&+(!%2BPW>!#(*!L8BPW>X(75;520$+(E!
/+$!@+$4+(*+2+!K&)0$5279#,!5(!*5+,+9!L$0)$'99!;',5+$2!'#8!+5(+9!,2067',25,67+(!dM2595+>
$#(),'&)0$5279#,!>!*+9!SK9+5,+('&)0$5279#,T!x/dPX-d et al.J!GHH`yE!/5+,+$!K&)0$5279#,!
&+7(2!,567!'(!*',!R+$7'&2+(!@0(!K9+5,+(!'(J!*5+! 5(!*+$!V')+!,5(*J!*+(!1Q$"+,2+(!_+)!"45>
! ! MATERIAL UND METHODEN
YH!
,67+(!57$+9!3+,2!#(*!+5(+$!:#22+$b#+&&+!"#!85(*+(E!U5+$8Q$!(#2"+(!,5+!5(*5$+12+!W099#(51'>
250(! 5(! :0$9! @0(! L7+$090(>9'$15+$2+(! L8'*+(E! K#8! *',! 90&+1#&'$+!Docking! '()+4+(*+2J!
85(*+2! *5+! '$2585"5+&&+! K9+5,+(10&0(5+! *',! %(+$)5+95(59#9! +5(+$! V5)'(*+(10(80$9'250(! 5(!
*+$! '125@+(! [',67+! *+,! L$02+5(,! 9522+&,! @+$,675+*+(+$! S/#82,M#$+(TE! U5+$;+5! 59525+$+(! *5+!
'$2585"5+&&+(! K9+5,+(! *',! R+$7'&2+(! @0(! $+'&+(! K9+5,+(E! O5+! 9'$15+$+(! (5+*+$+(+$)+25,67+!
V5)'(*+(10(80$9'250(+(!952!L7+$090(+(E!/5+!X(80$9'250(!*+,!'$2585"5+&&+(!L7+$090(M8'*+,!
45$*!*'((!5(!*'$'#8!80&)+(*+(!X2+$'250(+(!,0!90*585"5+$2J!*',,!(5+*+$+(+$)+25,67+!V5)'(*+(>
10(80$9'250(+(!952!+5(+$!7g7+$+(!_'7$,67+5(&5671+52!)+(+$5+$2!4+$*+(E!!!
:Q$! *5+! Docking>O2#*5+(! 952! LVK3[O! 4#$*+! *5+! LVK3[OcU%BLVL! Scoring>:#(1250(!
eLVK3[O>Scoref!xWdP< et al.J!CDDHy!@+$4+(*+2!e,5+7+!-&+567#()!CECfE!/5+,+!;+5(7'&2+2!+5>
(5)+![+$9+!'#,!*+$!c7+9>Score>X9M&+9+(25+$#()!5(!-dV/!45+!45(1+&';7A()5)+![+$9+!8Q$!
_',,+$,2088;$Q61+(;5(*#()+(! e CHOhbchhbhb fff JJ f! #(*! B+2'&&;5(*#()+(! e JJ coordmetmet ff !
chcoordmetchmet ff J fE! ^#*+9! +(27A&2! ,5+! 80&)+(*+! @5+$! +5)+(+! <+4+$2#(),109M0(+(2+(z! GE!
O2Q614+5,+,!&5(+'$+,!L02+(25'&!eLVL!{ Piecewise Linear Potentialf!x-+&7''$!et alEJ!CDDZyJ!CE!
+9M5$5,67+,!L02+(25'&! 8Q$!O674+$'209+!"#$!R+$75(*+$#()!@0(!5(2+$(+(!V5)'(*+(10&&5,50(+(J!
YE!%(+$)5+!*+$![0$,50(,,M'((#()!*+,!V5)'(*+(!#(*!]E!<5(*#(),+(+$)5+!*+$!5(2$'90&+1#&'$+(!
_',,+$,2088;$Q61+(!59!V5)'(*+(!e,5+7+!-&+567#()!CECfE!!
sitetorsclashchcoordmetchmet
coordmetmetCHOhbchhbhbPLPPLANTS
cffff
fffffffCHEMPLP 2.2
! !
%,! 4#$*+(! 80&)+(*+! L'$'9+2+$+5(,2+&&#()+(! 8Q$! *',! 90&+1#&'$+!Docking! @+$4+(*+2! e,5+7+!
[';+&&+!CEYfE!
Tabelle 2.3: Verwendete PLANTS-Parametereinstellungen. !
B#%+*.992%3.%!>)')(.*.';.1.#,-%2%3!#%!
>VW?MG!Q)92.+!
,+'$67!,+225(),! '60á'(2,! ZD!
! '60á+@'M! DECZ!
! '60á,5)9'! DECZ!
MATERIAL UND METHODEN
]D!
B#%+*.992%3.%!>)')(.*.';.1.#,-%2%3!#%!
>VW?MG!Q)92.+!
5(M#2! M$02+5(á85&+! %2BPWádL[XE90&C!
! &5)'(*á85&+! <5(*5()/<Y/áRCDDDá60$E90&C!
0#2M#2! 0#2M#2á*5$! ,DCZ'ZDMDCZ!
4$52+!,5()&+!90&C!85&+,! 4$52+á9#&25á90&C! G!
;5(*5()!,52+!*+85(5250(! ;5(*5(),52+á6+(2+$! >CEDIH!YCEFH]Y!GGECFCZ!
! ;5(*5(),52+á$'*5#,! G]!
,60$5()!8#(6250(! ,60$5()á8#(6250(! 67+9M&M!
6&#,2+$!'&)0$5279! 6&#,2+$á,2$#62#$+,! G!
! 6&#,2+$á$9,*! GED!!
!
JDRDP! B#%3.+.*1*.!".E.'*2%3+(.*-$0.%!CEFEYEG* PBO/>_+$2!
^#9!R+$)&+567!*+$!i#'&52A2!*+$! +5("+&(+(!Docking>!0*+$!Screening>%\M+$59+(2+!4#$*+!*+$!
PBO/>_+$2! ePBO/!{!Root-Mean-Square Deviationf!*+$!+5("+&(+(!O674+$'209+!7+$'()+>
"0)+(! xcdU%3!k! O[%P3<%P-J! GHFDyE! /5+,+$! )5;2! *+(! *#$67,67(522&567+(!K;,2'(*! "45>
,67+(!*+$!;+$+67(+2+(!V5)'(*+(M0,5250(!#(*!*+$!+\M+$59+(2+&&!)+8#(*+(+(!V5)'(*+(M0,5250(!
'(!e,5+7+!-&+567#()!CEYfE!!!
n
dRMSD i
iC
2.3
!
_0;+5!di!*5+!/588+$+("!"45,67+(!*+$!L0,5250(!*+,!O674+$'209,!i!*+,!V5)'(*+(!'#,!*+$!L/<!
#(*!*+$!L0,5250(!*+,!K209,!i!*+,!)+*0612+(!V5)'(*+(!*'$,2+&&+(E!/+$!K(2+5&!*+$!V5)'(*+(!952!
+5(+9!PBO/!1&+5(+$!0*+$!)&+567!C!h!45$*!'&,!10$$+12+!V5)'(*+(M0,+!)+4+$2+2E!!
!
! ! MATERIAL UND METHODEN
]G!
CEFEYEC* OM+'$9'(!W0$$+&'250(,10+885"5+(2!eP,f!
/5+! W0$$+&'250(,'('&N,+! "45,67+(! Scoring! #(*! *+9! PBO/>_+$2! 4#$*+(! ('67! OM+'$9'(!
*#$67)+8Q7$2!x_XOOV%PJ!GHDZyE!<+5!*5+,+$!K$2!K('&N,+!4#$*+!#(2+$,#672J!5(45+4+52!*5+!R'>
$5';&+(!Scoring>!#(*!RMSD>_+$2+!5(!<+"5+7#()!"#+5('(*+$!,2+7+(E!!
G
`G G
C
nn
dR
n
ii
s 2.4
!
/+$!75+$;+5!;+$+67(+2+!OM+'$9'(!W0$$+&'250(,10+885"5+(2!P,!1'((!"45,67+(!>G!#(*!G!@'$5+$+(!
e>G!{!5(@+$,+!W0$$+&'250(J!D!{!1+5(+!W0$$+&'250(J!G!{!W0$$+&'250(f!e,5+7+!-&+567#()!CE]fE!
!
CEFEYEY* K?c>_+$2!
?9!"#!Q;+$M$Q8+(J!0;!*5+!@+$4+(*+2+(!Docking>[00&,!+5(+!K($+567+$#()!@0(!'125@+(!O#;>
,2'("+(!';;5&*+(J!4#$*+!*5+!:&A67+!#(2+$!*+$!Pdc>W#$@+!ePdc!{!Receiver Operating Cha-
racteristicf!;+$+67(+2E!X(!*+$!Pdc>W#$@+!45$*!*5+!P'2+!*+$!S:'&,67>L0,525@+(T!e5('125@+!V5>
)'(*+(f!952!707+9!Docking-Score!"#$!P'2+!*+$!SP56725)>L0,525@+(T!e'125@+!V5)'(*+(f!'#8)+>
2$')+(E!/5+!;+$+67(+2+!:&A67+!#(2+$!*+$!Pdc>W#$@+!45$*!'&,!K?c>_+$2!eK?c!{!Area Under
Curvef! ;+"+567(+2! x/?/K et al.J! CDDGyE!/5+,+$!1'((!_+$2+! "45,67+(!(#&&! #(*!+5(,! '((+7>
9+(E!%5(!,67&+672+,!B0*+&&!4Q$*+!+5(+!8&'67+!W#$@+!5(!*+$!-+)+(*!*+$!_5(1+&7'&;5+$+(*+(!
952!+5(+9!K?c>_+$2!@0(!6'E!DJZ!&5+8+$(E!
!
CEFEYE]* K($+567+$#(),8'120$!e%:f!
/+$!Docking>K($+567+$#(),8'120$! e%:!{!Enrichment Factorf!)5;2!K#,1#(82!Q;+$!*5+!:A75)>
1+52J! 5((+$7'&;!@0(!Docking>%\M+$59+(2+(J!*5+!SP56725)>L0,525@+(T!)+)+(Q;+$!*+9!-+,'92>
*'2+(,'2"!7+$'#,"#85&2+$(J!59!R+$)&+567!"#!+5(+$!45&&1Q$&567+(!K#,4'7&!e,5+7+!-&+567#()!CEZf!
xcKRKOd[[d et al.J!CDDZyE!!!
total
totalxsampled
xsampled
HitsN
NHits
EF q
q
2.5
!
^#$!<+4+$2#()!*+$!K($+567+$#()+(!4#$*+(!*$+5!@+$,675+*+(+!K($+567+$#(),>:'120$+(!;+>
$+67(+2!e%:G!{!K($+567+$#()!5((+$7'&;!*+$!0;+$+(!Gq!*+$!/'2+(;'(1J!%:C!{!Cq!*+$!/'2+(>
;'(1J!Z!{!Zq!*+$!/'2+(;'(1fE!!
!
MATERIAL UND METHODEN
]C!
JDe! Docking 0.'!.'-)9*.%.%!G2;+*)%1.%!%),-!G2;+*'25*2'+2,-.%!B52!*+$!Docking>O0824'$+!-dV/!4#$*+!*5+!+$7'&2+(+!O#;,2'(";5;&5027+1J!'#,!*+(!O#;,2$#1>
2#$>!#(*!a7(&5671+52,,#67+(J!*5+!]]E]YD!O#;,2'("+(!+(275+&2J!'#8!+5(+9!V5(#\>Cluster!#(2+$>
,#672E!/5+!,0!)+40((+(+(!P07*'2+(!4#$*+(!*'((! 5(!+5(+$!L0,2>L$0"+,,5+$#()!4+52+$!'('&N>
,5+$2E!/5+!L0,2>L$0"+,,5+$#()!#98',,2+!*',!K#,,0$25+$+(!)+*0612+$!W0(80$9+$+J!*5+!;+,2599>
2+!W$52+$5+(!(5672!+$8Q&&+(J!+5(!Re-Scoring!#(*!*5+!'#209'25,5+$2+!%\2$'1250(!*+$!;+,2;+4+$2+>
2+(! O2$#12#$+(E! :Q$! *5+! %\2$'1250(! *+$! ;+,2;+4+$2+2+(! O2$#12#$+(! #(*! 57$+! d$5+(25+$#()+(J!
4#$*+!+5(![6&>;',5+$2+,!c'62@,>O1$5M2!@+$4+(*+2!45+!;+$+52,!@0(!<+N+$!;+,67$5+;+(!x<%j%PJ!
CDDIyE!!!
!/5+!'#,!*5+,+9!R+$8'7$+(!$+,#&25+$+(*+(!O2$#12#$+(!4#$*+(!*'((!'(7'(*!*+,!Scoring>_+$2+,!
'('&N,5+$2!#(*!;+4+$2+2E!/'@0(!4#$*+(!*5+!6'E!CDD!O2$#12#$+(!8Q$!p+*+!O2$#12#$1&',,+!952!*+(!
)Q(,25),2+(! <+4+$2#()+(J! 5(! *+$! <5(*+2',67+! @5,#+&&! ;+)#2'672+2E! /';+5! 4#$*+(! A7(&567+!
<5(*#(),90*5J!45+!,5+!8Q$!'(*+$+!X(75;520$+(!@0(!c/W,!5(!*+$!V52+$'2#$!;+,67$5+;+(!,5(*J!'&,!
B'r!+5(+$!)Q(,25)+(!X(2+$'1250(!952!*+$!%2cPWC!'()+,+7+(E!/5+!,0!)+8#(*+(+(!O#;,2'("+(!
4#$*+(!8Q$!;5067+95,67+!%\M+$59+(2+!'#,)+4A7&2E!
!
JDHO! Q#'*2.99.+!Screening!%),-!Y$*.%*#.99.%!B*IF=:!2%0!>&IF=:
/%-#;#*$'.%!:Q$!*5+! @5$2#+&&+!O#67+!('67!M02+(25+&&+(!%2BPW>!#(*!L8BPW>X(75;520$+(J!4#$*+!*5+!<5(>
*5()/<!/'2+(;'(1!5(!+5(+9!@5$2#+&&+(!Screening!)+M$Q82E!/5+,+!g88+(2&567!"#)A()5)+!/'2+(>
;'(1! +(27A&2! 6'E! CGEDDD! +\M+$59+(2+&&! ;+,25992+! <5(*#(),'885(52A2+(! 8Q$! L$02+5(>V5)'(*>
W09M&+\+! @0(! GGD! ^5+&M$02+5(+(! 952! @+$,675+*+(+(! X,080$9+(! #(*! B#2'(2+(! ,045+! 6'E!
GGEDDD!R+$;5(*#()+(E!:Q$!*',!@5$2#+&&+!Screening!4#$*+(!*5+!ZE]`G!W5(',+>X(75;520$+(!952!
*+$!Docking>O0824'$+!LVK3[O!p+4+5&,! 5(!*',!'125@+!^+(2$#9!*+$!%2BPW!#(*!*+$!L8BPW!
+5()+M',,2E! L$0! V5)'(*! 4#$*+(! p+4+5&,! "+7(!Docking>L0,+(! )+(+$5+$2E! /5+! <+4+$2#()! *+$!
L0,+(J! ,045+! *',! 85('&+! RankingJ! 4#$*+(! 952! *+$! LVK3[OcU%BLVL! Scoring>:#(1250(!
eLVK3[O>Scoref! *#$67)+8Q7$2E! /'('67! 4#$*+! 8Q$! *5+! "4+5! L$0"+(2! ;+,2+(! O2$#12#$+(! *+$!
85('&+(! /'2+(;'(1! *5@+$,+! B#$610! Assemblies! x<%BXO! k! B?PcWdJ! GHH`y! 9522+&,! *+$!
O0824'$+! L5M&5(+L5&02! )+(+$5+$2J! #9! )++5)(+2+!Hit-Cluster! 8Q$! *5+! %2BPW! #(*! *5+! L8BPW!
'#,"#4A7&+(E!!
!
! ! MATERIAL UND METHODEN
]Y!
JDHH! V#%.)'!/%*.'),*#$%!B%.'38![V/B]!".'.,-%2%3.%!!!K&&+! VX%>O59#&'250(+(! xKiRXO[! k! BKP%VX?OJ! CDDGy! 4#$*+(! 952! *+9! O0824'$+M'1+2!
Bd%J!#(2+$!R+$4+(*#()!*+,!BB::H]!W$'828+&*+,!xUKV-P%3J!GHH`y!*#$67)+8Q7$2J!A7(&567!
45+!;+$+52,!;+5!%()&5,7!et al.!;+,67$5+;+(!x%3-VXOUJ!CDDIyE!^#$!-+(+$5+$#()!*+$!%(,+9;&+,!
4#$*+(!B0&+1Q&*N('951,59#&'250(+(! @+$4+(*+2E! :Q$! '&&+! O59#&'250(+(!4#$*+! +5(+! ,M7A$5>
,67+!O59#&'250(,;0\!952!+5(+9!P'*5#,!@0(!GC!h!;+(#2"2!eK;;5&*#()!CEGfE!V5)'(*>!;"4E!V5>
)'(*>L$02+5(109M&+\+!4#$*+(! 5((+$7'&;!*+$!O59#&'250(,;0\!952!_',,+$90&+1Q&+(! 5(!Bulk-
dichte!,0&@'25,5+$2E!K&,!B0*+&&!8Q$!*',!_',,+$!4#$*+
MATERIAL UND METHODEN
]]!
!
/5+! O59#&'250(,,N,2+9+! 4#$*+(! 5(! +5(+9!9+7$,2#85)+(! L$0"+,,! +(+$)5+95(595+$2E! :Q$! *+(!
8$+5+(! ^#,2'(*! 4#$*+! +5(+! ^4+5M7',+(>B5(595+$#()! *#$67)+8Q7$2E! X(! *+$! +$,2+(! B5(595+>
$#(),M7',+! 4#$*+(! *5+! O674+$'209+! 952! 7'$90(5,67+(! PQ61,2+&&1$A82+(! @0(! CD! 16'&!
90& Gàh C!;+&+)2E!X(!*+$!"4+52+(!B5(595+$#(),M7',+!4#$*+(!*'((!'&&+!P+,2$51250(+(!'#8)+>
70;+(E!3'67+5('(*+$!4#$*+(!*5+!Methoden Steepest Descent! eO/f!xc?PPjJ!GH]]yJ!Conju-
gate Gradient! ec-f! xLKj3% et al.J! GHHCy!#(*!Truncated Newton! e[3f! xOcUVXcW!k!:d>
-%VOd3J! GHHCy! 5(! *+(! ;+5*+(! B5(595+$#(),M7',+(! '()+4+(*+2E! /5+! K;;$#67)$'*5+(2+(!
4#$*+(!p+4+5&,!'#8!!GEDDDJ!GDD!#(*!G!16'&!90& Gàh G!8+,2)+&+)2E!!
:Q$! *+(! )+;#(*+(+(!^#,2'(*!4#$*+(! *5+!-$'*5+(2+(! 5(! *+$![3!B+270*+! ';,67(5224+5,+! 5(!
+5(+$! R5+$M7',+(>B5(595+$#()! $+*#"5+$2E! X(! *+$! +$,2+(! B5(595+$#(),M7',+! 4#$*+(! '&&+!
O674+$'209+!85\5+$2!#(*!+5(![3!-$'*5+(2!@0(!GDD!16'&!90& Gàh G!@+$4+(*+2E!/'('67!4#$>
*+(! '&&+! O674+$'209+! *+,! P+"+M20$,! 952! 7'$90(5,67+(! PQ61,2+&&1$A82+(! @0(! ZDD! 16'&!
90& Gàh C!;+&+)2E!X(!+5(+9!4+52+$+(!B5(595+$#(),,67$522!4#$*+(!'&&+!P+"+M20$'209+J!*5+!5(!
+5(+9!P'*5#,! @0(! GZ!h! "#! *+9! ! V5)'(*+(! &5+)+(J! '&,! ,0)+('((2+! S<5(*+$+)50(T! *+85(5+$2E!
K&&+!O674+$'209+!*5+!,567! 5(!*5+,+$!P+)50(!;+85(*+(J!4#$*+(!952!+5(+$!W$'82!@0(!GDD!16'&!
90& Gàh C! 8+,2)+7'&2+(E!K#r+$7'&;! *5+,+$!P+)50(! ;+2$#)! *5+!P+,2$51250(,1$'82! 8Q$! O674+$>
'209+!ZDD!16'&!90& Gàh CE!:Q$!*5+,+!P+)50(!4#$*+!+5(![3!-$'*5+(2!@0(!GD!16'&!90& Gàh G!
@+$4+(*+2E!K;,67&5+r+(*!4#$*+(!'&&+!O674+$'209+J!,0407&!5(!*+$!<5(*+$+)50(!'&,!'#67!'#>
r+$7'&;!*5+,+$!P+)50(J!952!+5(+$!W$'82!@0(!CD!;"4E!GDD!16'&!90& Gàh C!8+,2)+7'&2+(E!U5+$8Q$!
4#$*+!+5(![3!-$'*5+(2!@0(!G!16'&!90& Gàh G!@+$4+(*+2E!!
/5+!B0&+1Q&*N('951,59#&'250(!4#$*+!59!1'(0(5,67+(!%(,+9;&+!59!K(,67&#,,!'(!*5+!"#@0$!
;+,67$5+;+(+(!%(+$)5+95(595+$#()+(! *#$67)+8Q7$2E! <+5! *5+,+9!%(,+9;&+!4+$*+(! *5+! [+5&>
67+('("'7&!e3fJ!*',!R0	+(!eRf!#(*!*5+![+9M+$'2#$!e[f!10(,2'(2!)+7'&2+(E!%,!4#$*+(!80&>
)+(*+!L'$'9+2+$+5(,2+&&#()+(!8Q$!*5+!B0&+1Q&*N('951!@+$4+(*+2!e,5+7+![';+&&+!CE]fE!
!
/5+! $+&'25@+(! <5(*#(),+(+$)5+(! 4#$*+(! '#8! <',5,! *+$! +$7'&2+(+(! [$'p+120$5+(J! 9522+&,! *+$!
VX%>B+270*+!)+9Ar!-&+567#()!GEG!!5(!Bd%!;+$+67(+2E!/5+!+\M+$59+(2+&&+(!W5>_+$2+!4#$>
*+(!5(!*5+!$+&'25@+(!<5(*#(),+(+$)5+(! -!9522+&,!-&+567#()!CE`!#9)+$+67(+2E!!
iKTRG &( 2.6 !
!
! ! MATERIAL UND METHODEN
]Z!
Tabelle 2.4: Verwendete Einstellungen der Moleküldynamiksimulation in MOE. !
>)')(.*.':!
;.1.#,-%2%3!#%!IZB!@.'*.! ".+,-'.#;2%3!
*2! DEDDGZ! 259+!,2+M!eM,f!
,'9M&+! GDDD! ,'9M&+!M+$50*!eM,f!
[D! CHFED! 5(525'&!@+&06525+,!2+9M+$'2#$+!
i[! GED! 2+9M+$'2#$+!$+,M0(,+!259+!eM,f!
iL! CED! M$+,,#$+!$+,M0(,+!259+!eM,f!
4'2+$c0(,2$'5(2,! â$5)5*â! xâ(0(+âJ!â;0(*,âJ!â$5)5*ây!
;0(*c0(,2$'5(2,! â&5)72â! xâ(0(+âJ!â&5)72âJ!â'&&ây!
60(,2$'5(2[0&! GED+>H! 20&+$'(6+!80$!70&0(0956!60(,2$'5(2,!
,'@+L0,5250(! D! ,'@+!M0,5250(,!20!*'2';',+!
,'@+R+&0652N! D! ,'@+!@+&06525+,!20!*'2';',+!
,'@+K66+&+$'250(! D! ,'@+!'66+&+$'250(,!20!*'2';',+!
+(,+9;&+! â3R[â! â3R[âJ!â3L[âJ!â3R%âJ!â3LUâ!
+(á,'9M&+! DEDDZ! 259+!80$!+(+$)N!,'9M&5()!
72! DED! 7+'25()!259+!eM,f!
+2! GEZ! âM$0*#6250(â!259+!80$!+b#5&5;$'250(!eM,f!
M$0*2! ZEZ! M$0*#6250(!259+!'2!+b#5&5;$5#9!2+9M+$'2#$+!eM,f!
62! DED! 600&5()!259+!eM,f!
7[! ZDED! 5(525'&!2+9M+$'2#$+!eWf!
+[! CHFED! +b#5&5;$5#9!2+9M+$'2#$+!eWf!
6[! CHFED! 600&5()!2'$)+2!2+9M+$'2#$+!eWf!
7L! GDGED! 5(525'&!M$+,,#$+!e1L'f!
+L! GDGED! +b#5&5;$5#9!M$+,,#$+!e1L'f!
6L! GDGED! 600&5()!2'$)+2!M$+,,#$+!e1L'f!
[á5(6! GDDED! 2+9M+$'2#$+!5(6$+9+(2!#M!20!+[!eWf!
MATERIAL UND METHODEN
]`!
JDHJ! K.%.'#.'2%3!.#%.'!5$(;#%)*$'#+,-.%!G2;+*)%10)*.%;)%5!?9!*5+!M02+(25+&&+(!V+52,2$#12#$+(!*+$!<+("595*'"0&>c'$;0(52$5&+!'#,!*+9!@5$2#+&&+(!Scree-
ning!4+52+$!"#!90*585"5+$+(J!4#$*+!+5(+!109;5('20$5,67+!O#;,2'(",2'("*'2+(;'(1!)+(+$5+$2E!
^#! *5+,+9!^4+61!4#$*+! +5(!c'62@,>O1$5M2! +$,2+&&2J! *',! "#(A67,2! O#;,252#+(2+(! '(! '$09'25>
,67+(!P5(),N,2+9+(!@0(!;+1'((2+(!W5(',+>X(75;520$+(! +$1+((2! ,045+! ';2$+((2E!/5+,+!:$')>
9+(2+! 4#$*+(! )+,0(*+$2! 5(! +5(+$! O#;,252#+(2+(*'2+(;'(1! ';&+)2E! /5+! ;+1'((2+(! W5(',+>
X(75;520$+(!,2'992+(!@0(!W5(',+>W'2'&0)+(!+5(5)+$!1099+$"5+&&+$!K(;5+2+$E!:$')9+(2+!952!
+5(+9!B0&+1#&'$)+45672!@0(!1&+5(+$!GFD!)o90&J!4#$*+(! 59!K(,67&#,,!'&,!O#;,252#+(2+(!'(!
*5+!-$#(*,2$#12#$!'(!*+(!L0,5250(+(!PG!;5,!PI!'()+8Q)2!eK;;5&*#()!CECfE!U5+$*#$67!4#$*+(!
5(,)+,'92!GCEYZ`!O2$#12#$+(!*+$!W&',,+!*+$!<+("595*'"0&>c'$;0(52$5&+!)+(+$5+$2E!/5+!67+95>
,67+! ^#)A()&5671+52! 4#$*+! 8Q$! *5+! -+(+$5+$#()! *+$! 109;5('20$5,67+(! O#;,2'(";5;&5027+1!
(5672!;+$Q61,56725)2E!!
%
%
$
%
T)
TE
TST2
TP
TC
TU
!
Abbildung 2.2: Substitutionsmöglichkeiten der Benzimidazol-Carbonitrile
^#$!-+(+$5+$#()!+5(+$!109;5('20$5,67+(!O#;,2'(";5;&5027+1J!4#$*+!'(!*+$!<+("595*'"0&>-$#(*,2$#12#$!'(!*+(!
L0,5250(+(!PG!;5,!PI!,#;,252#5+$2E!!
/5+!,0!+$7'&2+(+!109;5('20$5,67+!O#;,2'("*'2+(;'(1!4#$*+!*'((!*#$67!90&+1#&'$+,!Docking!
;+4+$2+2! #(*! 5(2+$+,,'(2+! V5)'(*+(! ,+&+125+$2E! :Q$! *5+! ,0! +$7'&2+(+(!R+$;5(*#()+(!4#$*+(!
Vg,&5671+52,@0$7+$,')+(! e,5+7+! K;,67(522! CEGYf! *#$67)+8Q7$2E! O#;,2'("+(! 952! +5(+9! )#2+(!
Vg,&5671+52,M$085&! 4#$*+(! ;+5;+7'&2+(! #(*! <5(*#(),+(+$)5+(! 9522+&,! *+$! VX%>B+270*+! ;+>
$+67(+2!e,5+7+!K;,67(522!CEGGfE!
!
JDHP! Vg+9#,-5.#*+6$'-.'+)3.%!^#$! R0$7+$,')+! @0(! Vg,&5671+52+(! +5("+&(+$! R+$;5(*#()+(! 4#$*+! *',! 27+$90*N('95,67+!
Vg,&5671+52,90*+&&! *+,! L$0)$'99M'1+2,! R0&,#$8t! x722Mzoo444E90&*5,60@+$NE609y! @+$4+(>
*+2E!R0&,#$8t!;+(#2"2!+5(!iOLP>B0*+&&!eiOLP!{!Quantitative Structure Property Relation-
shipfE!/',!B0*+&&!;',5+$2!'#8!6'E!GEGDD!67+95,67+(!O2$#12#$+(E!R0&,#$8t!,')2!*5+!27+$90*N>
! ! MATERIAL UND METHODEN
]I!
('95,67+!Vg,&5671+52!&0)O!5(!_',,+$!;+5!CHF!W!@0$7+$J!40;+5!O!5(!90&o&!'()+)+;+(!45$*E!/+$!
O2'(*'$*8+7&+$!*+$!R0$7+$,')+!45$*!@0(!*+(!K#20$+(!952!+24'!ÅDJI!&0)'$52795,67+(!%5(7+5>
2+(! '()+)+;+(! x722Mzoo444E90&*5,60@+$NE609yE! O0952! 1'((! *5+!B+270*+! '&,! :5&2+$#(),9+>
270*+!8Q$!*5+!@5$2#+&&+!O#;,2'("*'2+(;'(1!+5()+,+2"2!4+$*+(E!
!
JDHS! In vitro!I.++2%3.%!/5+!?(2+$,#67#()+(!*+$!9522+&,!O#;,2$#12#$>!#(*!a7(&5671+52,,#67+! 5*+(2585"5+$2+(!O#;,2'(>
"+(!'#8!;50&0)5,67+!K125@52A2J!4#$*+(!@0(!B'$�'!V#�,'!O#Ä$+"!:+$Ä(*+"!eX(2+$@+2!X((0@'250(!
-9;UJ!O674';+(7+59f!*#$67)+8Q7$2E!/';+5!4#$*+!*5+!X(75;5250(!*+$!1'2'&N25,67+(!K125@52A2!
*+$! %("N9+! 5(! K(4+,+(7+52! *+$! @+$,675+*+(+(! O#;,2'("+(! )+9+,,+(E! :Q$! *5+! B+,,#()+(!
4#$*+! *+$! STR-FRET Screening Express KitT! @0(! B0&+6#&'$! /+@56+,! @+$4+(*+2!
x722Mzoo444EB0&+6#&'$*+@56+,E609yE! :Q$! *5+! R+$,#67+! 4#$*+(! *5+! %2cPWCJ! %2cjcY'! #(*!
*',!PX3-d>L$02+5(!x:%P<j et al.J!GHHHy!*+,!W$'&&+(8$0,67,!Xenopus laevis!5(!+5(+9!Esche-
richia coli!%\M$+,,50(,,N,2+9!$+109;5('(2!+\M$595+$2!#(*!'#8)+$+5(5)2E!/',!L$02+5(!PX3-d!
;5(*+2!#(,M+"585,67!'(!@+$,675+*+(+!c/W,!#(*!5,2!45+!%2cjcY'!+5(!K125@'20$!*+$!%2cPWCE!
/5+!'125@5+$2+!%2cPWC!4#$*+!('67!U+$,2+&&+$@0$)';+(!5(!*+9
ERGEBNISSE
]F!
P!B'3.;%#++.!
!
PDH! "#$#%&$'()*#+,-.!G.f2.%1)%)98+.!6$%!4A=+!2%0!7859#%.%!6$%!!E. tenella
R0(! *+$!E. tenella! -+(09,+b#+("! 4#$*+! 59! :$Q7p'7$! CDDI! +5(! +$,2+$! %(24#$8! eDraft As-
semblyf!8+$25)!)+,2+&&2J!*+$! p+*067!(067!(5672!@0&&,2A(*5)!'((025+$2!4#$*+E!/5+!O+b#+("*'2+(!
4#$*+(! '9! O'()+$! X(,252#2J! 59! P'79+(! +5(+,! E. tenella! -+(09>O+b#+("5+$#(),M$0p+12+,J!
)+(+$5+$2! #(*! ,5(*! '#8! +5(+9! g88+(2&567! "#)A()&567+(! :[L>O+$@+$! 8$+5! @+$8Q);'$! 75(2+$&+)2!
x82Mzoo82ME,'()+$E'6E#1oM#;oM'270)+(,o%59+$5'o2+(+&&'yE! :Q$! *5+,+! <VKO[>K('&N,+(! 4#$*+(!
;+$+52,! ;+1'((2+! c/W>! ,045+! ^N1&5(>A7(&567+! L$02+5(,+b#+("+(! @0(! d$)'(5,9+(! *+$!Api-
complexaJ!'&,!K;8$')+,+b#+("+(!@+$4+(*+2E!
!
PDHDH! 4A=:-$($9$3.!>'$*.#%.!#%!E. tenella
^#!<+)5((!*5+,+$!K$;+52!4'$!(#$!+5(+!c/W!@0(!E. tenella!;+1'((2!e%2cPWCn!cPW!{!CDK-
Related KinasefE! %2cPWC! 4#$*+! ;+$+52,! @0(! +5(+$! 100M+$5+$+(*+(! K$;+52,)$#MM+! 1&0(5+$2J!
+\M$595+$2!#(*!;5067+95,67!67'$'12+$5,5+$2!xWX33KXP/ et al.J!CDD]yE!O5+!)5&2!'&,!+5(!7090>
&0)+,! L$02+5(! *+,! 7#9'(+(!c/WC! L$02+5(,! eU,c/WCf! xWX33KXP/ et al.J! CDD]yE!K#8! *+$!
K95(0,A#$+,+b#+("+;+(+!;+,2+72!+5(+!a7(&5671+52!@0(!`Fq!"#$!U,c/WC!,045+!"#!*+$!7#>
9'(+(!c/WG! eU,c/WGfE!^#!'(*+$+(!c/W>A7(&567+(!L$02+5(+(!*+$!ApicomplexaJ!45+!;+5>
,M5+&,4+5,+! *+$! L8LWZ! eL8LWZ! {! Plasmodium falciparum! L$02+5(! W5(',+! Zf! xPdOO>
BKc/d3KV/ et al.J! GHH]yJ! ;+,2+72! +5(+! O+b#+("A7(&5671+52! "45,67+(! IZq! #(*! FDq!
xWX33KXP/ et al.J!CDD]yE!!
K(*+$,!'&,!c/W,!@0(!R+$2+;$'2+(J!*5+!Q;+$!*5+!)+,'92+!O+b#+("&A()+!7067!10(,+$@5+$2!@0$>
&5+)+(!xBKV?B<P%O!k!<KP<KcX/J!CDDZyJ!@'$55+$2!*5+!K95(0,A#$+';80&)+!*+$!c/W!O+>
b#+("+(! *+$!Apicomplexa,! 5(,;+,0(*+$+! '9!c>[+$95(#,E!<+1'((2+!c/W,!*+$!Apicomplexa!
4+5,+(!O+b#+("5*+(252A2+(!@0(!4+(5)+$!'&,!]Dq!5(!*+(!GDD!c>2+$95('&+(!K95(0,A#$+(!'#8E!
3+;+(! *+(! 2+5&4+5,+! )$0r+(! O+b#+("#(2+$,675+*+(! '9! c>[+$95(#,J! ,5(*! '#67! *5+! ^N1&5(>
<5(*+9025@+! eLO[KXP%>B025@f! 5(! 57$+$! O+b#+("';80&)+! ,+7$! @+$,675+*+(E! ^#*+9! "+5)+(!
*5+,+! L$02+5(+! 082! 3>2+$95('&+! #(*! c>2+$95('&+! R+$&A()+$#()+(E! %5(+! L$02+5(9025@'('&N,+!
'#8!<',5,!*+$!B025@*'2+(;'(1+(!L8'9!#(*!LPdOX[%!x<K[%BK3 et al.J!CDD]n!U?Vd et al.J!
CDD`y! +$)';J! *',,!4+52+$+! 8Q$!W5(',+(! 2NM5,67+!O+b#+("9025@+! 5(! '&&+(!cPW!L$02+5(+(!*+$!
! ! ERGEBNISSE
]H!
Apicomplexa! +(27'&2+(! ,5(*E! O5+! ;+,52"+(! 9+7$+$+! ,2'$1! 10(,+$@5+$2+! P+)50(+(! 59! '125@+(!
^+(2$#9E!U5+$"#!"A7&+(!*5+!Hinge>P+)50(J!*+$![>Loop!#(*!+5(!-&N65(>$+567+,!O+b#+("9025@!
e->Loopf! e,5+7+!K;;5&*#()!GEH! #(*!K;;5&*#()!YECfE!U5+$;+5! "+5)2+! ,567J! *',,! *+$!-$'*! *+$!
W0(,+$@5+$#()!@0(!K95(0,A#$+(!5(!8#(1250(+&&+(!<+$+567+(!*+$!%("N9+!4+52'#,!,2A$1+$!'#,>
)+M$A)2!5,2E!%5(+!"+(2$'&+!P0&&+!,M5+&2!"#*+9!*5+!10(,+$@5+$2+!K95(0,A#$+!/G]]!e3#99+$5+>
$#()!('67!*+$!K95(0,A#$+,+b#+("!*+$!%2cPWCfJ!*5+! 8Q$!*5+!1'2'&N25,67+!K125@52A2!@0(!+(2>
,67+5*+(*+$!<+*+#2#()!5,2E!O5+!5,2!<+,2'(*2+5&!*+$!1'2'&N25,67+(!^+(2$#9,!WYCJ!%ZD!#(*!/G]]!
e%2cPWCf!x<PK3/%3!k![dd^%J!GHHHyE!/5+,+!K95(0,A#$+(!&5+)+(!59!9#&25M&+(!Alignment!
*+$!cPW>L$02+5(,+b#+("+(!*+$!Apicomplexa!'(!5*+(25,67+(!L0,5250(+(!@0$E!!!
^#$! X*+(2585"5+$#()! @0(! 4+52+$+(! cPW>O+b#+("+(! 5(! E. tenellaJ! 4#$*+(! O#67+(! 952! *+9!
<VKO[}>L$0)$'99!5(!*+$!)+(095,67+(!O+b#+("*'2+(;'(1!@0(!E. tenella!*+,!O'()+$!X(,25>
2#2+,!x722Mzoo444E,'()+$E'6E#1y!*#$67)+8Q7$2E!:Q$!*5+,+!<VKO[>K('&N,+(!4#$*+(!*5+!,5+;+(!
;+$+52,! ;+1'((2+(! cPW,! @0(!P. falciparum! ! >! L8LWZJ! L8LW`J! L8BPWJ! L8cPWGJ! L8cPWYJ!
L8cPW]!#(*!L8cPWZ!x<PKccUX>PXcKP/ et al.J!CDDDn!/d%PX- et al.J!GHHZn!/d%PX- et
al.J!CDDCn!/d%PX-J!CDD]n!VX et al.J!GHH`n!PdOO>BKc/d3KV/ et al.J!GHH]y!l!'&,!K;8$')+>
,+b#+("+(!@+$4+(*+2E!U5+$;+5!10((2+(!(+;+(!*+$!;+$+52,!;+1'((2+(!%2cPWC!xWX33KXP/ et
al.J! CDD]yJ! *5+! 7090&0)! "#! L8LWZ! 5,2J! *$+5! #(;+1'((2+! cPW,! @0(!E. tenella! @0$7+$)+,')2!
4+$*+(E!O5+!4#$*+(!5(!K(&+7(#()!'(!*5+!3'9+(!*+$!+(2,M$+67+(*+(!L$02+5(+!@0(!P. falcipa-
rum!eL8BPWJ!L8cPWG!#(*!L8cPWYf!%2BPWJ!%2cPWG!#(*!%2cPWY!)+('((2E!/5+!a7(&567>
1+52,;+"5+7#()+(!*+$!c/W>A7(&567+(!L$02+5(+!@0(!P. falciparumJ!"#!*+(!(+#!5*+(2585"5+$2+(!
L$02+5(+(!5(!E. tenellaJ!,5(*!5(![';+&&+!YEG!#(*!K;;5&*#()!YEG!*'$)+,2+&&2E!!!
V+*5)&567!8Q$!*5+!%2BPW!10((2+!+5(+!@0&&,2A(*5)+!O+b#+("!5*+(2585"5+$2!4+$*+(J!4A7$+(*!8Q$!
%2cPWG!#(*!%2cPWY!(#$![+5&,+b#+("+(!)+8#(*+(!4#$*+(!e,5+7+![';+&&+!YEG!#(*!K;;5&*#()!
YEGfE!/5+!/'2+(;'(1,#67+(!952!*+(!K;8$')+,+b#+("+(!L8LW`J!L8cPW]!#(*!L8cPWZ!+$)';+(!
1+5(+!,5)(5851'(2+(![$+88+$E!K#8)$#(*!*+$!59!R+$)&+567!"#!'(*+$+(!c/W>O+b#+("+(!*+$!Api-
complexa! ,+7$!)+$5()+(!O+b#+("A7(&5671+52J! ,5(*!*5+,+!%$)+;(5,,+!(5672!#()+4g7(&567E!^#>
*+9!5,2!'&&)+9+5(!;+1'((2J!*',,!*5+!W5(',+(!L8LW`J!L8cPW]!,045+!L8cPWZ!A7(&567+!X*+(25>
2A2,4+$2+!"#!*+(!c/W,J!'&,!'#67!"#!*+(!BKL!W5(',+(!eBKLW!{!Mitogen-Activated Protein
Kinasef!'#84+5,+(!x/d%PX- et al.J!CDDCn!_KP/ et al.J!CDD]yJ!4+,7'&;!,5+!'#67!'&,!UN;$5*>
M$02+5(+! @0$7+$)+,')2! 4+$*+(! x-%j%P et al.J! CDDZyE! /5+,+! ['2,'67+! "+5)2! ,567! '#67! 5(!
<VKO[L!K('&N,+(!*+$!L8cPW]!#(*!*+$!L8cPWZJ!*5+!,0407&!O+b#+("9025@+!@0(!c/W,J!'&,!
'#67!@0(!BKLW,!'#84+5,+(!x_KP/ et al.J!CDD]yE!/5+!L8LW`!(5992!75+$;+5!+5(+!O0(*+$,2+&>
ERGEBNISSE
ZD!
&#()! +5(J! *'! ,5+! 088+(,5672&567! *5+! +5("5)+!c/W>A7(&567+! O+b#+("! 5(!P. falciparum! 5,2J! *5+!
1+5(!^N1&5(;5(*+9025@!;+,52"2! xcU%3 et al.J! CDD`yJ! ,0952! 1g((2+! ,5+! '#67! '&,! +5(+!^N1&5(>
#(';7A()5)+! c/W! '()+,+7+(! 4+$*+(J! 4',! ;5067+95,67! ;+,2A25)2! 4#$*+! x<PKccUX>
PXcKP/ et al.J!CDDDyE!!
!!
Abbildung 3.1: Schematischer Sequenzvergleich von CDK-ähnlichen Proteinen von P. falciparum und E. tenella.
3+;+(! *+(! ;+$+52,! ;+1'((2+(! O+b#+("+(! @0(!P. falciparum! #(*! *+$! %2cPWC! @0(!E. tenellaJ! 4#$*+(!9522+&,!
<VKO[>O#67+(! *$+5! (+#+!c/W>A7(&567+! O+b#+("+(! 8Q$!E. tenella! 5*+(2585"5+$2E! /5+!K;;5&*#()! "+5)2! *5+! *$+5!
@0$7+$)+,')2+(!L$02+5(,+b#+("+(!,045+!%2cPWCE!/5+!c/W>A7(&567+(!O+b#+("+(!,5(*!p+4+5&,!'&,!L''$+!"#!*+(!
7090&0)+(!O+b#+("+(! 5(!P. falciparum!';)+;5&*+2E!:Q$!*5+!%2cPWG!#(*!*5+!%2cPWY!10((2+(! p+*067!(#$!#(>
@0&&,2A(*5)+!L$02+5(,+b#+("+(!@0$7+$)+,')2!4+$*+(E!/5+!#(@0&&,2A(*5)+(!O+b#+("';,67(522+!,5(*!952!+5(+9!S|T!
9'$15+$2E!/',! 67'$'12+$5,25,67+!^N1&5(><5(*+9025@! "+5)2!R'$5'250(+(! 5(! *+(!c/W>A7(&567+(!L$02+5(,+b#+("+(!
e@50&+22fE!_+52+$+! 67'$'12+$5,25,67+! O+b#+("9025@+! ,5(*z!->Loop! e)+&;fJ!Hinge! e)$Q(f! #(*!K125@5+$#(),>Loop!
e$02fE!!
8! :G"HHJKL!
8! :G"HIJKL!
FB_!J0)1#6#S#!
GUG&A!
L#:K&?!
=! ?CC!
?DF!
8! :85RNNKL!
8! :95HHJKL!
GU<K&!
L#<K&!
=! ;?@!
@F_!J0)1#6#S#!
8! :I<H"NM
`8! :`GRH"NKL!
GU:K&=!
L#:K&=!
@BD!;CE!=! B=D!
BF_!J0)1#6#S#!
8! :I&HQJKL!
8! :`G&HIJKL!
GU:K&;!
L#:K&;!
BCB!;DC!=! =;@=!
@F_!J0)1#6#X!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
!
=! ;=@!
=!
=!
DD!
?@;!
=!
! ! ERGEBNISSE
ZG!
!!!
Abbildung 3.2: Multiples Sequenzalignment von HsCDK2 und CDK-Homologen von E. tenella.
>U+&56+,!#(*! >:'&2;&A22+$!,5(*!#(2+$!*+(!O+b#+("+(!'&,!$0,'!<&g61+J!;"4E!'&,!)$Q(+!L8+5&+!*'$)+,2+&&2E!^#!*+(!
67'$'12+$5,25,67+(!O+b#+("9025@+(!"A7&+(!LO[KXP%>B025@!e@50&+22fJ!->Loop!e)+&;fJ!Hinge>$+)50(!e)$Q(fJ!/:->
B025@!e;&'#f!#(*!K125@5+$#(),>Loop!e$02fE!/',!^N1&5(><5(*+9025@!eLO[KXP%>B025@f!@'$55+$2!5(!*+$!O+b#+("';>
80&)+!5(!cPW>A7(&567+(!L$02+5(+(!*+$!ApicomplexaE!/5+!K95(0,A#$+(!*+,!1'2'&N25,67+!^+(2$#9,!WYCJ!%ZD!#(*!
/G]]! e%2cPWC!3#99+$5+$#()f! ,5(*!*#$67!S|T!9'$15+$2E!K95(0,A#$+(!*5+!*#$67!ShT!1+(("+567(+(!*5+!K[L>
<5(*#(),,2+&&+(E!/5+!L70,M70$N&5+$#(),,2+&&+(![G]J!jGZ!#(*
ERGEBNISSE
ZC!
Tabelle 3.1: Vergleich der bekannten CDK-ähnlichen Sequenzen von P. falciparum und E. tenella sowie den
Aktivatoren.
O#67+(!952!*+9!<VKO[>K&)0$5279#,!+$9g)&5672+(!*5+!X*+(25851'250(!@0(!7090&0)+(!c/W>L$02+5(+(!e%2BPWJ!
%2cPWG!#(*!%2cPWYfE!%2cPWG!,045+!%2cPWY!10((2+(!;5,7+$!(#$!#(@0&&,2A(*5)!5*+(2585"5+$2!4+$*+(E!:Q$!*5+!
[+5&,+b#+("+(! ,5(*!*5+! '()+)+;+(+(! X*+(252A2+(!952! +5(+9!S|T!9'$15+$2E!K&,!K125@'20$+(!*+$!%2cPWC!*5+(+(!
cN6&5(!K!xWX33KXP/ et al.J!CDD]y!,045+!PX3-d!#(*!%2cjcY'!xO?äP%^!:%P3ä3/%^J!#(M#;&5"5+$2+!/'>
2+(J!X(2+$@+2!X((0@'250(!-9;UJ!O674';+(7+59yE!
!
YEGEGEG* %2BPW!
%2BPW!4+5,2!+5(+!)$0r+!3>2+$95('&+!R+$&A()+$#()!@0(!]Y!K95(0,A#$+(!e;+"+567(+2!59!:0&>
)+(*+(!'&,!%2BPWá&'()f!'#8E!R5+&+!'(*+$+!7090&0)+!L$02+5(,+b#+("+(J!5((+$7'&;!*+,!O2'9>
9+,! *+$!ApicomplexaJ! ;+,52"+(! +5(+(! *+#2&567! 1Q$"+$+(!3>[+$95(#,E! %5(+! A7(&567+J! p+*067!
(067! &A()+$+!%\2+(,50(! 5,2! 8Q$!*5+!['cPWY!'#,!Theileria annulata!;+1'((2! xWX33KXP/ et
al.J! CDDGyE! X9! :'&&+! *+$! ['cPWY! ;+5(7'&2+2! *5+,+! 3>2+$95('&+! O+b#+("! +5(! 9#29'r&567+,!
W+$(>V01'&5,5+$#(),,5)('&! e3VO! {!Nuclear Localization Signalf! xWX33KXP/ et al.J! CDDGn!
B%X%P!k!<Vd<%VJ!GHHDyE!:Q$!*5+!%2BPWá&'()!10((2+!952!*+9!O0824'$+>Tool!M$+*5623VO!
1+5(!3VO>W0(,+(,#,9025@!@0$7+$)+,')2!4+$*+(E!^#*+9!4#$*+(!5(!*5+,+$!R+$&A()+$#()!1+5>
Plasmodium falciparum! Eimeria tenella!
K.%.!i!>'$*.#%! W5*#6)*$'! >'$*.#%!/0.%*#*<*!12!
P. falciparum >'$*.#%
W5*#6)*$'!
L8LWZ!!xPdOO>BKc/d3KV/ et al.J!
GHH]y!
6N6UJ!MCZJ!L8cjcGJ!
L8cjcYJ6N6KJ!PX3-d!xV%!PdcU
et al.J!CDDDn!B%PcW} et al.J!
CDDYy!
%2cPWC!xWX33KXP/ et al.J!
CDD]y!`Iq!
cN6&5(!K!xWX33KXP/ et al.J!CDD]yJ!PX3-dJ!%2cjcY'!xO?äP%^!
:%P3ä3/%^J!#(M#;&5"5+$2+!/'2+(y!
L8LW`!!x<PKccUX>PXcKP/ et al.J!CDDDy!
cN6&5(>5(*+M+(*+(2!x/d%PX- et al.J!
CDDCy!! ! !
L8BPW!!xVX et al.J!GHH`y!
6N6UJ!L8cjcG!xV%!PdcU et al.J!CDDDn!_K[%PO et
al.J!CDDDy!
EtMRK ! ]Cq! !
L8cPW>G!x/d%PX- et al.J!GHHZy! ! EtCRK1! I`q|! !
L8cPW>Y!x/d%PX- et al.J!CDDCy! ! EtCRK3! ]`q|! !
L86$1>]!!x/d%PX- et al.J!CDDCy! ! ! ! !
L86$1>Z!x_KP/ et al.J!CDD]y! ! ! ! !
! ! ERGEBNISSE
ZY!
(+!4+52+$+(J! ;+1'((2+(!O+b#+(">B025@+!)+8#(*+(J!4',!@+$9#2+(! &A,,2J! *',,! +5(!O+b#+("5+>
$#(),8+7&+$! @0$&5+)2E! /5+! 3#1&+025*,+b#+("'('&N,+! +$)';! 59! V+,+$'79+(! tG! +5(! SKc-Ö>
[$5M&+2!!'(!L0,5250(!GYH!*+$!%29$1E!:'&&,!5((+$7'&;!*+,!SKc-T!+5(!O+b#+("5+$#(),8+7&+$!@0$>
&5+)2!#(*!9'(!,2'22*+,,+(!+5(!SK[-T!'((5992J!+$7A&2!9'(!+5(!W0"'1>W0(,+(,#,>B025@E!/',!
@+$9+5(2&567+!O2'$260*0(!*+$!%2BPWá&'()>O+b#+("!5,2!75()+)+(!(5672!5(!+5(+9!W0"'1>B025@!
+(27'&2+(E!K#,!*5+,+9!-$#(*!4#$*+!8Q$!'&&+!4+52+$+(!K$;+52+(!*5+!1Q$"+$+!3#1&+025*,+b#+("!
'()+(099+(E!/5+!@0$7+$)+,')2+!%2BPW!L$02+5(,+b#+("!$+,#&25+$2+!,0! 5(!+5(+$!@+$)&+567;'>
$+(! VA()+! #(*! K95(0,A#$+';80&)+! "#! '(*+$+(! 7090&0)+(! BPW>O+b#+("+(! 5((+$7'&;! *+$!
Apicomplexa!e,5+7+!K;;5&*#()!YEYfE!/5+!%2BPW>L$02+5(,+b#+("!5,2!"#!]`q!5*+(25,67!952!*+$!
@+$4'(*2+(! P. falciparum! L8BPW>L$02+5(,+b#+("E! /5+! K('&N,+! *+,! ,M+"585,67+(! ^N1&5(>
<5(*+9025@+,!+$)';!*5+!K95(0,A#$+(';80&)+!SU:[[XP%T!8Q$!*5+!%2BPWE!/5+,+,!B025@! 5,2!
8Q$! '&&+!BPW>A7(&567+(! L$02+5(+! 4+52)+7+(*! 10(,+$@5+$2! e,5+7+! [';+&&+! YECfE! K&&+! '(*+$+(!
W5(',+>B025@+!45+!->LoopJ!Hinge>P+)50(J![>Loop!#(*!/:->B025@!10((2+(!+;+(8'&&,!5(!*+$!
%2BPW!5*+(2585"5+$2!4+$*+(E!!!
K#,!*+$!V52+$'2#$!5,2J!59!-+)+(,'2"!"#!c/W>L$02+5(+(!@0(!OA#)+$(J!(#$!4+(5)!Q;+$!*5+!:#(1>
250(!7090&0)+$!BPW!L$02+5(+!*+$!Apicomplexa!;+1'((2!xcK3/?PX et al.J!CDDIn!cU%3 et
al.J!CDD`n!WX33KXP/ et al.J!CDDGyE!<+5,M5+&,4+5,+!,67+5(2!L8BPW!+5(+!'(*+$+!P+)#&'250(,>
8#(1250(!"#!;+,52"+(J!'&,!*5+!A7(&567+(!OA#)+$M$02+5(+!xcU%3 et al.J!CDD`n!WKV/XOJ!GHHHyE!
/5+! L8BPW! ;+,52"2J! 59! R+$)&+567! "#! c/W>L$02+5(+(! @0(! OA#)+$(J! *5+! 7g67,2+! X*+(252A2!
eYFqf! "#9! 7#9'(+(! c/WI! L$02+5(! eU,c/WIn! x)5ã]ZDCI]YyfE! U,c/WI! ;+,52"2! +5(+! *#'&+!
:#(1250(! x:XOU%PJ!CDDZyE!%5(+$,+52,! 5,2!U,c/WI! 5(!*+$
ERGEBNISSE
Z]!
cKW!8Q$!*5+!L8BPWJ!952!L8LWZ!'&,!+5(!9g)&567+,!O#;,2$'2J!;5,7+$!;5067+95,67!(5672!;+,2A>
25)2!xcU%3 et al.J!CDD`yE!!
!
Abbildung 3.3: Sequenzalignment von MRK-ähnlichen Proteinen der Apicomplexa sowie HsCDK7.
K&5)(5+$2! 4#$*+(! *5+! BPW! L$02+5(,+b#+("+(! @0(! Cryptosporidium hominis! ec7BPWfJ! Theileria annulata!
e['BPWfJ!Plasmodium falciparum! eL8BPWfJ!Plasmodium yoelii yoelii! eLNNBPWf! ,045+! *5+! 7#9'(+!c/WI!
L$02+5(,+b#+("!eU,c/WIfE!/5+!456725)+(!O+b#+("9025@+!,5(*!59!Alignment!9'$15+$2E!K95(0,A#$+(J!*5+!5(!'&&+(!
L$02+5(,+b#+("+(!10(,+$@5+$2!@0$&5+)+(J!,5(*!;&'#!#(2+$&+)2E!-+&;!#(2+$&+)2+!K95(0,A#$+(!,5(*!5(!8Q(8!O+b#+(>
"+(J!,045+!)$'#!#(2+$&+)2+!K95(0,A#$+(!,5(*!5(!@5+$!O+b#+("+(!10(,+$@5+$2E!!
?9! "#! #(2+$,#67+(J! 0;! *',! %2BPW!L$02+5(! +;+(8'&&,! '(! *+$! [$'(1,1$5M250(,$+)#&'250(! *+$!
P3K>L0&N9+$',+!XX!;+2+5&5)2!5,2J!4#$*+!*5+!E. tenella!-+(09,+b#+("!('67!+5(+$!"#!L8BK[G!
;"4E!+5(+$!"#!U,BK[G!7090&0)+(!O+b#+("!*#$67,#672E!%5(+!<VKO[}>O#67+!952!L8BK[!5(!
*+$!E. tenella!-+(09,+b#+("!5*+(2585"5+$2+!+5(+!7090&0)+!%2BK[GE!/5+,!&A,,2!@+$9#2+(J!*',,!
*',!L$02+5(!%2BPW!+;+(8'&&,!'(!*+$
! ! ERGEBNISSE
ZZ!
2$'(,&0"5+$2! 4+$*+(! 9Q,,+(E! /5+! 7#9'(+! c/WI! O+b#+("! 4+5,2! 59! -+)+(,'2"! "#$! %2BPW!
O+b#+("!+5(!3VO>O5)('&!'#8E!!!
/5+!:$')+J!0;!%2BPW!+;+(8'&&,!+5(+!cKW!:#(1250(!;+,52"2J!1'((!(5672!'&&+5(!*#$67!O+b#+(">
@+$)&+567+!@0$7+$)+,')2!4+$*+(!#(*!,0&&2+!*'7+$!;5067+95,67!#(2+$,#672!4+$*+(E!=+*067!)5;2!
+,! +5(5)+! U5(4+5,+! *5+! )+)+(! +5(+! ,0&67+! :#(1250(! ,M$+67+(z! eXf! %,! 10((2+! 1+5(! cKW>
<5(*+9025@! 5(!*+(!M02+(25+&&+(!O#;,2$'2+(!%2cPWC!xWX33KXP/ et al.J!CDD]y!#(*!%2cPWY!
5*+(2585"5+$2!4+$*+(E!eXXf!/5+!L70,M70$N&5+$#()!*+,!K125@5+$#(),>Loops!*+$!%2cPWC!#(*!*+$!
L8LWZ!1'((!'#67!*#$67!K#20M70,M70$N&5+$#()!+$80&)+(E!:Q$!*5+!L8LWZ!4#$*+!;5,&'()!(067!
(5672!;+,2A25)2J!0;!*5+,+!K#20M70,M70$N&5+$#()!*5+!+5("5)+!K125@5+$#(),9+270*+!5,2!xcU%3 et
al.J!CDD`n!WX33KXP/ et al.J!CDD]yE!
Tabelle 3.2: MRK-ähnliche Proteine der Apicomplexa sowie deren Zyklin-Bindedomänen. !
Z'3)%#+(2+! ?)(.!!?4"/!!
G.f2.%1:/A!
7859#%:
"#%0.0$(<%.!
/0.%*#*<*!12!
>&IF=!!^j_
Plasmodium falciparum L8BPW! }LáDDGY]I]C`! NFVLLRE GDD!
Plasmodium berghei L;BPW! }LáI``YDC! NFLILRE HG!
Plasmodium knowlesi L1BPW! }LáDDCCZFIFZ! NFVLLRE FF!
Plasmodium vivax L@BPW! }LáDDG`G]]ZZ! NFVLLRE FI!
Plasmodium yoelii yoelii LNNBPW }LáICZZZZ! NFLILRE FZ!
Crypotsporidium hominis c7BPW! }Lá``ZFHH! HFTVLRE ZD!
Crypotsporidium muris c9BPW! }LáDDCG]CDIG! HFTVLRE ]H!
Crypotsporidium parvum cMBPW! }LáDDGYFFY`I! HFTVLRE ZG!
Babesia bovis <;BPW! }LáDDG`GDH]Z! HFTALRE ]I!
Theileria annulata ['BPW! }Lá`IF`DY! HFTTLRE ]H!
Theileria parva [MBPW! }LáI``YDC! HFTTLRE ]F!
Toxoplasma gondii [)BPW! }LáDDCY`ZIDH! HFTTVRE YH!
Eimeria tenella %2BPW! >! HFTTIRE ]Y!!
ERGEBNISSE
Z`!
YEGEGEC* %2cPWG!
_5+! ;+$+52,! 5(! K;,67(522! YEGEG! ;+,67$5+;+(J! 10((2+! *5+! %2cPWG! ;5,&'()! (#$! #(@0&&,2A(*5)!
@0$7+$)+,')2!4+$*+(E!:Q$!;+5*+![+$95(5!*+$!%2cPWG!K95(0,A#$+,+b#+("+(!&')+(!1+5(+!/'>
2+(!5(!*+(!/'2+(;'(1+(!@0$E!/5+!%2cPWG!L$02+5(2+5&,+b#+("!;+,2+72!'#,!HH!K95(0,A#$+(E!O5+!
;+5(7'&2+2! *',! ^N1&5(;5(*+>B025@! SLR[OVP%TJ! 4+&67+,! 5((+$7'&;! *+$! 7090&0)+(! cPWG>
L$02+5(,+b#+("+(!*+$!Apicomplexa!4+52)+7+(*!10(,+$@5+$2!@0$&5+)2!e,5+7+![';+&&+!YEYfE!K(*+>
$+! ,M+"585,67+! W5(',+>O+b#+("9025@+! 10((2+(! '#8)$#(*! *+$! #(@0&&,2A(*5)+(! O+b#+("! (5672!
5*+(2585"5+$2!4+$*+(E!/5+!;505(80$9'25,67!5*+(2585"5+$2+!%2cPWG>[+5&,+b#+("!5,2!"#!I`q!5*+(>
25,67!"#$!;+1'((2+(!L8cPWG!L$02+5(>O+b#+("E!/5+!W0(,+$@5+$#()!'9!3>!#(*!'9!c>[+$95(#,!
*+$!cPWG>7090&0)+(!L$02+5(+!5,2!59!R+$)&+567!"#9!9522&+$+(!O+b#+("';,67(522!*+#2&567!)+>
$5()+$J!40*#$67!*5+!X*+(2585"5+$#()!*+$!2+$95('&+(!O+b#+("';,67(522+!+$,674+$2!5,2E!!
Tabelle 3.3: : CRK1-ähnliche Proteine der Apicomplexa sowie deren Zyklin-Bindedomänen. !
Z'3)%#+(2+! ?)(.!!?4"/!!
G.f2.%1:/A!
7859#%:
"#%0.0$(<%.!
/0.%*#*<*!12!
>&4F=H!^j_
Plasmodium falciparum L8cPWG! }LáDDGYZG]F` AMTSLRE GDD!
Plasmodium chabaudi
chabaudi L66cPWG! }LáI]CHZ`! AITSLRE I`!
Plasmodium knowlesi L1cPWG! }LáDDCCZFCY] AITSLRE FG!
Plasmodium vivax L@cPWG! }LáDDG`GZDFF AITSLRE II!
Plasmodium yoelii yoelii LNNcPWG! }LáIYGYGY! AITSLRE II!
Babesia bovis <;cPWG! }LáDDG`DHH`G PVSYLRE ]H!
Theileria annulata ['cPWG! }LáHZYCDC! PLTYLRE ZD!
Theileria parva [McPWG! }LáI`]GF`! PLTNLRE Z]!
Toxoplasma gondii [)cPWG! }LáDDCYIDCHG PVTSLRE YH!
Eimeria tenella %2cPWG! >! PVTSLRE I`!
!
! ! ERGEBNISSE
ZI!
YEGEGEY* %2cPWY!
%2cPWY!5,2!*+$!U,c/WG!eY`q!O+b#+("5*+(252A2f!,045+!*+$!L8cPWY!A7(&567!e]`q!O+b#+(">
5*+(252A2fE!/5+,+!K95(0,A#$+,+b#+("!10((2+!+;+(8'&&,!(#$!#(@0&&,2A(*5)!@0$7+$)+,')2!4+$*+(E!
_A7$+(*! 8Q$! *+(!3>[+$95(#,! +5(! O2'$260*0(! )+8#(*+(!4#$*+J! 10((2+! 8Q$! *+(! c>[+$95(#,!
1+5(!O2'$260*0(!5*+(2585"5+$2!4+$*+(E!/5+!%2cPWY!L$02+5(,+b#+("!;+,2+72!'#,!C]Y!K95(0,A#>
$+(E! /',! ^N1&5(><5(*+9025@! 5,2! 8Q$! '&&+! cPWY>A7(&567+(! L$02+5(+! 4+52)+7+(*! 10(,+$@5+$2!
e,5+7+![';+&&+!YE]fE!/5+!@+$;&+5;+(*+(!W5(',+>,M+"585,67+(!O+b#+("9025@+!10((2+(!+;+(8'&&,!
5(!*+$!%2cPWY!L$02+5(,+b#+("!5*+(2585"5+$2!4+$*+(E!%5(5)+!*+$!7090&0)+(!cPWY!O+b#+("+(!
'(*+$+$!Apicomplexa!;+,52"+(!+5(+!R+$&A()+$#()!*+,!3>[+$95(#,J!45+!+$!;+5,M5+&,4+5,+!;+5!
*+$!L8cPWY!L$02+5(,+b#+("!@0$&5+)2E!/5+!3>2+$95('&+(!R+$&A()+$#()+(!4+5,+(!+5(+!)+$5()+!
O+b#+("5*+(252A2!"#+5('(*+$!'#8E!/5+,+!3>2+$95('&+!R+$&A()+$#()!10((2+!p+*067!(5672!8Q$!*5+!
%2cPWY!L$02+5(,+b#+("!5*+(2585"5+$2!4+$*+(E!!!
%5(!4+52+$+$!,5)(5851'(2+$!?(2+$,675+*!*+$!%2cPWY!#(*!L8cPWY!85(*+2!,567!59!9522&+$+(!<+>
$+567! *+$! L$02+5(,+b#+("J! "45,67+(! *+(!/09A(+(! *+$! c>U+&5\! #(*! *+$!Hinge>P+)50(E! /5+!
L8cPWY!L$02+5(,+b#+("!4+5,2!'(!*5+,+$!O2+&&+!+5(+!,+7$!)$0r+!X(,+$250(!@0(!+24'!CDD!K95(0>
,A#$+(! e,5+7+! K;;5&*#()! YE]f! '#8E! a7(&567+! X(,+$250(+(! 4+$*+(! '#67! ;+5! cMcPWY! #(*!
[McPWY!)+8#(*+(J!p+*067!(5672!;+5!U,c/WG!eK;;5&*#()!YE]fE!K#,!*5+,+9!-$#(*!,67+5(2!*5+!
O+b#+("109M0,5250(!@0(!%2cPWY!A7(&567+$!"#$!U,c/WG!'&,!"#!*+$!L8cPWY!"#!,+5(E!!!
/5+!K95(0,A#$+(![7$G]!#(*![N$GZ!*+$!U,c/WG!,5(*!'(!*+$!X('125@5+$#()!*+,!c/WGo^N1&5(!
W09M&+\+,!;+2+5&5)2!#(*!67'$'12+$5,25,67!8Q$!c/WG!A7(&567+!L$02+5(+E!/5+!X('125@5+$#()!+$>
80&)2!*#$67!L70,70$N&5+$#()E!_A7$+(*![N$GZ!;+5!'&&+(!cPWY>A7(&567+(!O+b#+("+(!*+$!Api-
complexa! 10(,+$@5+$2! @0$&5+)2J! 5,2! [7$G]! 5(! *+$! %2cPWY! O+b#+("! *#$67! +5(! K&'(5(! '#,)+>
2'#,672! e,5+7+! K;;5&*#()! YE]fE! /5+,! &A,,2! @+$9#2+(J! *',,! *5+! X('125@5+$#()! *5+,+,! L$02+5(,!
9g)&567+$4+5,+! '#8! +5(+! '(*+$+! K$2! $+)#&5+$2! 45$*E! 3+;+(! *+$! X('125@5+$#(),$+)#&'250(! 5,2!
'#67!*5+!)+('#+!:#(1250(!*+$!cMcPWY!,045+!'&&+$!cPWY!7090&0)+(!L$02+5(+!;5,&'()!(5672!
;+1'((2!xcK3/?PX et al.J!CDDIyE!
ERGEBNISSE
ZF!
!
Abbildung 3.4: Sequenzalignment von CRK3-ähnlichen Proteinen der Apicomplexa sowie HsCDK1.
X9!Alignment! ,5(*!456725)+!O+b#+("9025@+!9'$15+$2E!K95(0,A#$+(J! *5+! 5(! '&&+(!L$02+5(,+b#+("+(!10(,+$@5+$2!
@0$&5+)+(J!,5(*!;&'#!#(2+$&+)2E!-+&;!#(2+$&+)2+!K95(0,A#$+(!,5(*!5(!8Q(8!O+b#+("+(J!)$'#!#(2+$&+)2+!K95(0,A#>
$+(!5(!@5+$!O+b#+("+(!10(,+$@5+$2E!
DFG-Motif
M*
T-Loop
Hinge
C-Helix
M*G-Loop
M*
! ! ERGEBNISSE
ZH!
Tabelle 3.4: CRK3-ähnliche Proteine der Apicomplexa sowie deren Zyklin-Bindedomänen. !
Z'3)%#+(2+! ?)(.!!?4"/!!
G.f2.%1:/A!
7859#%:
"#%0.0$(<%.!
/0.%*#*<*!12!
>&4F=P!^j_
Plasmodium falciparum L8cPWY! }LáDDGYZG]`G! AKTYIRE GDD!
Plasmodium berghei L;cPWY! }Lá`IICFF! AKTYIRE IY!
Plasmodium chabaudi
chabaudi L66cPWY! }LáI]ZCHG! AKTYIRE IY!
Plasmodium knowlesi L1cPWY! }LáDDCCZFCDY! AKTYIRE ZD!
Plasmodium vivax L@cPWY! }LáDDG`GZDY]! AKTYIRE `H!
Plasmodium yoelii yoelii LNNcPWY! }LáIC]CI]! AKTYIRE YH!
Cryptosporidium hominis c7cPWY! }Lá``FG``! SKTAIRE ]Z!
Cryptosporidium muris c9cPWY! }LáDDCG]DIGC! SKTAIRE ]]!
Cryptosporidium parvum cMcPWY! }LáDDGYFFYGC! SKTAIRE ]]!
Babesia bovis <;cPWY! }LáDDG`DFF]I! PKSAIRE ]`!
Theileria annulata ['cPWY! }LáHZ]F]D! PKNAIRE ZD!
Theileria parva [McPWY! }LáI`YGZF! PKNAIRE ZD!
Toxoplasma gondii [)cPWY! }LáDDCY`IC`D! PRTAIRE ]Z!
Eimeria tenella %2cPWY! >! PKTAIRE ]`!!
!
PDHDJ! 7859#%.!6$%!E. tenella!
/5+!O#67+!('67!^N1&5(+(!952!a7(&5671+52+(!"#!;+$+52,!;+1'((2+(!^N1&5(+(!'#,!P. falciparumJ!
4#$*+(!9522+&,!<VKO[>O#67+(! 5(! +5(+$!-+(09>/'2+(;'(1!@0(!E. tenella! *#$67)+8Q7$2E!%,!
10((2+(!@5+$!O+b#+("+(! 5*+(2585"5+$2J! ,045+!*5+!+(2,M$+67+(*+(!L$02+5(,+b#+("+(! e%2cjcGJ!
%2cjcYKJ!%2cjcY<!#(*!%2cjc]n!K;;5&*#()!YEZ!,045+!K;;5&*#()!YE`f!@0$7+$)+,')2!4+$>
*+(E!/5+,+!L$02+5(,+b#+("+(!;+,52"+(!a7(&5671+52+(!"#!*+(!^N1&5(+(!L8cjcGJ!L8cjcY!#(*!
L8cjc]!@0(!P. falciparum!e,5+7+!K;;5&*#()!YEZ!#(*!K;;5&*#()!YE`fE!X(!+5(+9!K125@5+$#(),>
Assay!952!%2cjcY'!,045+!%2cPWC!10((2+!B'$�'!VE!O#Ä$+"!:+$(Ä(*+"!eX(2+$@+2!X((0@'250(!
ERGEBNISSE
`D!
-9;UJ!O674';+(7+59f!"+5)+(J!*',,!%2cjcY'!+5(!)++5)(+2+$!K125@'20$!*+$!%2cPWC!5,2E!%2>
cjcY'!5,2!"#!CCq!"#!L8cjcY!#(*!"#!GIq!"#!U,cjcK!5*+(25,67E!!
!
Abbildung 3.5: Schematischer Vergleich von Zyklin-ähnlichen Proteinen von P. falciparum und E. tenella.
B52!*+(!;+1'((2+(!^N1&5(+(!@0(!P. falciparumJ!10((2+(!9522+&,!<VKO[>O#67+(!@5+$!^N1&5(>A7(&567+!O+b#+("+(!
@0(!E. tenella!5*+(2585"5+$2!4+$*+(E!/5+!)+8#(*+(+(!^N1&5(>A7(&567+(!O+b#+("+(!,5(*!p+4+5&,!'&,!L''$+!"#!57$+(!
7090&0)+(!O+b#+("+(!5(!P. falciparum!*'$)+,2+&&2E!:Q$!%2cjcG!#(*!%2cjcY;!e952!+5(+9!S|T!9'$15+$2f!10((>
2+(!p+*067!(#$!#(@0&&,2A(*5)+!L$02+5(,+b#+("+(!7+$)+&+52+2!4+$*+(E!/',!67'$'12+$5,25,67+!^N1&5(><0\!B025@!5,2!
;&'#!9'$15+$2E!!!
/5+! O#67+!952! *+$! K;8$')+,+b#+("! L8cjcG! @0(!P. falciparum! +$)';! 1+5(+! A7(&567+(! O+>
b#+("+(! 5(! E. tenellaE! K#8)$#(*! *+$! )+$5()+(! O+b#+("5*+(252A2! *+$! ^N1&5(+! *+$! +5("+&(+(!
W&',,+(!"#+5('(*+$J!4#$*+!*5+!O#67+!952!*+$!"#!L8cjcG!7090&0)+(!O+b#+("!'#,!T. gondii!
e^N1&5(!Uf!*#$67)+8Q7$2E!/'!*5+!d$)'(5,9+(!T. gondii! #(*!E. tenella! ;+5*+!*+$!W&',,+!*+$!
Coccidia '()+7g$+(J! 1'((!952!W0(,+$@5+$#()! *+$! O+b#+("+(! )+$+67(+2!4+$*+(E!K#,! *5+,+$!
O#67+! $+,#&25+$2+! +5(+J! 5(! *+$!V52+$'2#$! ;5,&'()!(5672! ;+,67$5+;+(+J!^N1&5(>A7(&567+!O+b#+("!
@0(!E. tenella!e,5+7+!K;;5&*#()!YEZn!%2cjcGfE!/+$!10$$+12+!V+,+$'79+(!4#$*+!9522+&,!M''$>
8! :
8! :
GUV:Q:;!
L#:Q!:;$!
=! ??D!
??_!J0)1#6#S#!;EE!
@F_!J0)1#6#S#!
L#:Q:;+!
=! ?F?!8! :
8! :
GU:Q:@!
L#:Q:@!
?=_!J0)1#6#S#!@?E!
;?F!
8! :
8! :`
=@_!J0)1#6#S#!
GU:Q:=!
L#:Q:=!
=!
=! =@=!
=!
= =;C
=!
`8! :`
! ! ERGEBNISSE
`G!
4+5,+9!Alignment!;+,25992!#(*!*5+! $+,#&25+$+(*+!3#1&+025*,+b#+("! 5(!+5(+!L$02+5(,+b#+("!
2$'(,&'25+$2E!/'"#!4#$*+!*+$!K&)0$5279#,!transeq!@+$4+(*+2!xPXc%!et alEJ!CDDDyE!/5+!2$'(,&'>
25+$2+! L$02+5(>O+b#+("! ;+,52"2! GGZ! K95(0,A#$+(! #(*! 5,2! "#! G]q! 952! *+$! L$02+5(,+b#+("!
L8cjcG!'#,!P. falciparum 5*+(25,67 e,5+7+!K;;5&*#()!YEZfE!%5(+!InterproScan>K('&N,+!;+,2A>
25)2!*',!R0$&5+)+(!+5(+$!^N1&5(>A7(&567+(!/09A(+E!/5+!2$'(,&'25+$2+!L$02+5(,+b#+("!%2cjcG!
5,2!p+*067!,0407&!'9!3>[+$95(#,J!'&,!'#67!'9!c>[+$95(#,!*+$!K95(0,A#$+,+b#+("!#(@0&&>
,2A(*5)E!!!
!
Abbildung 3.6: Sequenzalignment von Zyklin-ähnlichen Proteinen von E. tenella.
<VKO[>O#67+(!+$)';+(!@5+$!^N1&5(>A7(&567+!O+b#+("+(!5(!E. tenellaE!U5+$!*'$)+,2+&&2!,5(*!*5+!@0$7+$)+,')2+(!
L$02+5(,+b#+("+(E!:Q$!*5+!%2cjcG!#(*!*5+!%2cjcY;!10((2+(!p+*067!(#$!#(@0&&,2A(*5)+!O+b#+("+(!5*+(2585"5+$2!
4+$*+(E!!<&'#!#(2+$&+)2+!K95(0,A#$+(!,5(*!5(!@5+$!O+b#+("+(J!)+&;!#(2+$&+)2+!K95(0,A#$+(!5(!*$+5!O+b#+("+(!
,045+!)$'#!#(2+$&+)2+!K95(0,A#$+(!5(!"4+5!O+b#+("+(!10(,+$@5+$2E!!
/#$67!O+b#+(",#67+!952! *+$!K;8$')+,+b#+("!L8cjcCJ! 10((2+! 1+5(+!O+b#+("! 5(!E. tenella!
+$9522+&2! 4+$*+(J! p+*067! 5*+(2585"5+$2+! *5+! O+b#+("! L8cjcY! "4+5! A7(&567+! O+b#+("+(E! /5+!
ERGEBNISSE
`C!
O+b#+(",#67+!952!*+$!K;8$')+,+b#+("!L8cjc]!+$)';!+5(+!(+#+!O+b#+("! 5(!E. tenellaE!:Q$!
*5+! 9522+&,! *+$! K;8$')+,+b#+("+(! L8cjcY! #(*! L8cjc]! )+8#(*+(+(! O+b#+("+(! e%26N6Y'J!
%26N6Y;!#(*!%26N6]fJ!10((2+!*#$67!+5(+(!*5$+12+(!O+b#+("@+$)&+567!*+$!10$$+12+!V+,+$'79+(!
5*+(2585"5+$2!4+$*+(E!:Q$!*5+!O+b#+("+(!@0(!%26N6Y'!#(*!%26N6]!10((2+(!1+5(+!X(2$0(,!5(!*+$!
)+(095,67+(!O+b#+("!5*+(2585"5+$2!4+$*+(J!4',!p+*067!*',!R0$7'(*+(,+5(!@0(!X(2$0(,!(5672!
@0&&,2A(*5)! '#,,67&5+r2E! K;;5&*#()! YE`! "+5)2! +5(!9#&25M&+,!Alignment! @0(! ^N1&5(>A7(&567+(!
L$02+5(+(!@0(!E. tenellaE!
!
PDJ! ZY*#(#.'2%3!2%0!W%'.#,-.'2%3!6$%!B*4F=J:/%-#;#*$'.%!X(!*+$!K$;+52!@0(!<+N+$! x<%j%PJ!CDDIy!4#$*+(! 5(!+5(+9!5(525'&+(J!@5$2#+&&+(!Screening!'(!
%2cPWC! ;+$+52,! +5(5)+! Hit-Compounds! 5*+(2585"5+$2E! /5+,+! 4#$*+(! '(,67&5+r+(*! *#$67! in
vitro![+,2#()!;+,2A25)2E!^5+&!*5+,+$!K$;+52!4'$!+,J!*5+!5*+(2585"5+$2+(!Hits!"#!Hit-Cluster!'("#>
$+567+$(J!,045+!*5+!L02+("!*5+,+$!Compounds!*#$67!)+"5+&2+!dM2595+$#()!"#!+$7g7+(E!
!
PDJDH! G2;+*'25*2':!2%0!d-%9#,-5.#*++2,-.%!^#$! K($+567+$#()! M02+(25+&&+$! X(75;520$+(! *+$! %2cPWC! 4#$*+(! O#;,2$#12#$>! #(*! a7(&567>
1+52,,#67+(!*#$67)+8Q7$2E!K&,!K#,)'(),,#;,2'("+(!*5+(2+(!+&8!59!@5$2#+&&+(!Screening!)+8#(>
*+(+!X(75;520$+(J!*5+!+5(+(!XcZD>_+$2!@0(!1&+5(+$!GDD!ÜB!59!%("N9>Assay!"+5)2+(E!:Q$!*5+!
O2$#12#$,#67+(!4#$*+(!"#(A67,2!)++5)(+2+!O#;,2$#12#$+(!'#,)+4A7&2!e,5+7+![';+&&+!YEZfE!/5+!
K#,4'7&! 8'(*! '(7'(*! *+$!-$#(*,2$#12#$! (Scaffoldf! ,2'22J! *5+! 59!90&+1#&'$+(!Docking! +5(+!
67'$'12+$5,25,67+!_',,+$,2088;$Q61+(>X(2+$'1250(! 952! *+$!Hinge>P+)50(! *+$! %2cPWC! "+5)2+!
eWarheadf!e,5+7+!K;,67(522!GEZEYfE!X(,)+,'92!10((2+(!,0!]]EYZG!O#;,2'("+(!'#,!*+$!X(2+$@+2!
c7+956'&!/'2';',+!eXc/f!,+&+125+$2!4+$*+(J!4+&67+!*5+!)+4Q(,672+(!O#;,2$#12#$+(!"+5)2+(E!!!
/5+! a7(&5671+52,,#67+(!4#$*+(!9522+&,! +5(+,! L5M&5(+L5&02!WorkflowsJ! 45+! 5(! K;,67(522! CE`!
;+,67$5+;+(J! *#$67)+8Q7$2E! %,! 10((2+(! CEYYG! A7(&567+! R+$;5(*#()+(!952! +5(+9! ['(59020>
a7(&5671+52,10+885"5+(2+(! @0(! 95(*+,2+(,! IDq! "#$! K#,)'(),,2$#12#$! 5*+(2585"5+$2! 4+$*+(E!
/5+!3#2"#()!+5(+$!a7(&5671+52,,#67+!7'2!)+)+(Q;+$!*+$!O#;,2$#12#$,#67+!*+(!R0$2+5&J! *',,!
O#;,2'("+(!)+8#(*+(!4+$*+(J!*5+!@0(!57$+$!O#;,2$#12#$!"#$
! ! ERGEBNISSE
`Y!
Tabelle 3.5: Substruktursuchen zur Anreicherung von EtCRK2-Inhibitoren. !
G*'25*2'59)++.! G2;+*'25*2'!W%1)-9!3.&2%0.%.'!G*'25:
*2'.%!#%!0.'!/4A!
<'$;52#$'2+! % %
Q
$ $
&
!GDE]HC!
<+("595*'"0&+!%
%
%!GDE`G`!
3'M720&'62'9+!%
$
&
!YH]!
d\5(*0&+!%
&$ !
GIEZGY!
P70*'(5(+! %
Q
&
Q
$ !YEDF]!
!
B52! *+(! 75+$! )+"+5)2+(! O2$#12#$1&',,+(! 4#$*+(! O#;,2$#12#$,#67+(! 5(! *+$! X(2+$@+2! c7+956'&! /'2';',+! eXc/f!
*#$67)+8Q7$2E!/5+!K#,4'7&!)++5)(+2+$!O2$#12#$1&',,+(!8'(*!'(7'(*!*+$!-$#(*,2$#12#$!,2'22J!*5+!952!*+$!Hinge>
P+)50(!Q;+$!_',,+$,2088;$Q61+(!5(2+$')5+$2E!!
!
PDJDJ! I$9.529)'.+!Docking!12'!W2+E)-9!6#'*2.99.'!Hits!
/5+! 9522+&,! O#;,2$#12#$>! ;"4E! a7(&5671+52,,#67+(! )+8#(*+(+(! R+$;5(*#()+(! 4#$*+(! 5(!
O2$#12#$>Cluster!"#,'99+()+8',,2E!/5+!K("'7&!*5+,+$!O2$#12#$+(!4'$!p+*067!"#!)$0rJ!#9!*5+>
,+!O#;,2'("+(!59! in vitro
ERGEBNISSE
`]!
,2Q61! eK1"+M20$>! 0*+$!/0(0$'209f! +5(+$! V5)'(*+(M0,+! 59!P'*5#,! *5+,+$!W#)+&J! 4#$*+! *5+!
V5)'(*+(>L0,+!;+5;+7'&2+(E!!
R0(! *+(! L0,+(J! *5+! +5(+(! 4'7$,67+5(&567+(! <5(*#(),90*#,! $+M$A,+(25+$+(J! 4#$*+! 9522+&,!
+5(+$!^4+5,2#8+(><+4+$2#()!xjdd3!k!_%VOUJ!CDD]y!+5(+!P'()80&)+!+$,2+&&2E!K(7'(*!*5+>
,+$! P'()80&)+J! 4#$*+! *5+! O2$#12#$'#,4'7&! 801#,,5+$2E! /5+! 0M259'&+! ^4+5,2#8+(><+4+$2#()!
4#$*+!;+$+52,!@0(!<+N+$!;+,67$5+;+(! x<%j%PJ!CDDIyE!/';+5!1'9+(!"4+5!@+$,675+*+(+!<+>
4+$2#(),8#(1250(+(! "#9! %5(,'2"E! X(! +5(+$! +$,2+(! <+4+$2#()! e='5(>ScoringfunktionJ! x=KX3J!
GHH`yf!4#$*+!"#(A67,2!*5+!L0,+!+5(+$!O2$#12#$!'#,)+4A7&2J!*5+!*+$!4'7$,67+5(&567,2+(!;50'1>
25@+(!W0(80$9'250(!+(2,M$5672E!X(!+5(+$!"4+52+(!<+4+$2#()!e->ScoreJ!x=d3%O et al.J!GHHIyf!
4#$*+(! *5+! @+$;&+5;+(*+(! L0,+(! @+$,675+*+(+$! O2$#12#$+(! 952+5('(*+$! @+$)&567+(E! U5+$;+5!
4#$*+!*5+!+(*)Q&25)+!P'()80&)+!+$9522+&2E!!!
?9!_5$1,2088+!"#!85(*+(J!*5+!,+&+125@!*5+!%2cPWC!5(75;5+$+(J!,0&&2+(!*5+,+!*5+!(+)'25@!)+&'>
*+(+!K95(0,A#$+!-&#FF!'*$+,,5+$+(E!K(!*+$!'('&0)+(!L0,5250(!;+,52"2!*5+!c/WY!*+,!U#7(,!
*5+!M0,525@!)+&'*+(+!K95(0,A#$+!VN,FH!e,5+7+!K;;5&*#()!YEIfE!V5)'(*+(!952!+5(+$!8#(1250(+&>
&+(!-$#MM+J! *5+! *5+,+!V'*#(),#91+7$! '*$+,,5+$+(J! ,0&&2+(! *'*#$67! ,+&+125@! '(! *5+!%2cPWC!
;5(*+(E! :Q$! *5+! O+&+1250(! +(2,M$+67+(*+$! O2$#12#$+(! 4#$*+! 45+*+$#9! +5(! X(2+$'1250(,85&2+$!
@+$4+(*+2!x<%j%PJ!CDDIyE!!
/#$67!*',!DockingJ!*5+!C>O2#8+(><+4+$2#()!#(*!*5+!X(2+$'1250(,85&2+$!10((2+(!'#,!*+(!*#$67!
O2$#12#$,#67+(!$+,#&25+$+(*+(!O#;,2'("+(!e5(,)+,'92!]`E`FCfJ!`EZFH!%5("+&,#;,2'("+(!,+&+1>
25+$2!4+$*+(!e,5+7+
! ! ERGEBNISSE
`Z!
!
Abbildung 3.7: Primär- und Tertiärstruktur der EtCRK2 und deren ATP-Bindetasche.
Kf!O+b#+("'&5)(9+(2!"45,67+(!U,c/WC!#(*!%2cPWC!,045+!*+$!c/WY!*+,!U#7(,!e-)c/WYfE!/5+,+,!Align-
ment! *5+(2+! '&,! <',5,! 8Q$! *5+! %$,2+&&#()! *+,! U090&0)5+90*+&&,E! -$'#! 75(2+$&+)2+! K95(0,A#$+(! ,5)('&5,5+$+(!
GDDq5)+!s;+$+5(,2599#()E!P02!e10(,+$@5+$2+f!#(*!)+&;!eK95(0,A#$+('#,2'#,67f!+5()+8A$;2+!K95(0,A#$+(!*+$!
K[L><5(*+2',67+E!K95(0,A#$+(J! *5+!952!<#67,2';+(! )+1+(("+567(+2! ,5(*J! )+7g$+(! "#! *+$!K*+(5(;5(*+$+)50(!
eKfJ!*+$!P5;0,+;5(*+$+)50(!ePf!#(*!*+$!L70,M7'2;5(*+$+)50(!eLfE!K95(0,A#$+(J!*5+!"#!*+(!;+5*+(!O+&+125@52A2,>
2',67+(!)+7g$+(J!,5(*!952!$g95,67+(!^588+$(!X!;"4E!XX!)+1+(("+567(+2E!
<f!U090&0)5+90*+&&!*+$!%2cPWC!@0(!E. tenella!e;&'#f!5(!P5;;0(>/'$,2+&&#()!x<%j%PJ!CDDIyE!^#9!R+$)&+567!
4#$*+!*5+!7#9'(+
ERGEBNISSE
``!
Tabelle 3.6: Screening nach potentiellen EtCRK2-Inhibitoren. !
G.9.5*#$%++,-'#**.!0.'!Hit:W%'.#,-.'2%3! W%1)-9!0.'!Q.';#%02%3.%!
O2$#12#$,#67+(! ]`E`FC!
Docking ]`ED`H!
3'67M$0"+,,5+$#()! `EZFH!
Compound>K#,4'7&! ZDZ!
<+,2+&&2+!O#;,2'("+(! CFH!
<5067+95,67+![+,2,! GZH!
K125@+!R+$;5(*#()+(! `!!
U5+$!,5(*!*5+!+5("+&(+(!O+&+1250(,,67$522+!*+$!Hit>K($+567+$#()!*'$)+,2+&&2E!!
!
PDJDP! In vitro!M.+*2%3!0.'!)2+3.E<-9*.%!G2;+*)%1.%!
R0(!*+(!;+,2+&&2+(!O#;,2'("+(!e5(,)+,'92!CFHf!*+$!W&',,+(!*+$!<+("595*'"0&+J!3'M720&'62'>
9+!#(*!P70*'(5(+!10((2+(!GZH!2'2,A67&567!)+&5+8+$2!4+$*+(E!/5+!K125@52A2!*5+,+$!O#;,2'("+(!
4#$*+! 5(! +5(+9! %2cPWC>%("N92+,2! ;+,25992E! :Q$! *5+! B+,,#()+(! 4#$*+! *+$! TR-FRET
Screening Express Kit @0(!B0&+6#&'$!/+@56+,!x722Mzoo444EB0&+6#&'$*+@56+,E609y!@+$4+(*+2!
e,5+7+!K;,67(522!CEG]fE!O+67,!*+$!GZH!O#;,2'("+(!45+,+(!+5(+(!XcZD>_+$2!1&+5(+$!'&,!GDD!ÜB!
'#8! e,5+7+! [';+&&+! YEIfE! :Q$! '&&+! '125@+(! O#;,2'("+(! 4#$*+(! *5+! Vg,&5671+52+(! 5(! _',,+$!
(+M7+&09+2$5,67!;+,25992! eV5'(+!_'&2+$J! X(2+$@+2! X((0@'250(!-9;UJ!O674';+(7+59fE!O#;>
,2'("+(!*5+!+5(+!Vg,&5671+52!)$gr+$!'&,!ZDD!ÜB!"+5)2+(J!4#$*+(!9522+&,!VcoBO>!#(*!3BP>
B+,,#()+(!+5(+$!i#'&52A2,10(2$0&&+!#(2+$"0)+(!eO2+8'(!/+$,67#9!#(*!c7$5,25'(!O670$(J! X(>
2+$@+2!X((0@'250(!-9;UJ!O674';+(7+59fE!!!
?9!'#8"#1&A$+(!0;!+,!,567!;+5!*+(!O#;,2'("+(! 2'2,A67&567!#9!K[L>109M+2525@+! X(75;520$+(!
7'(*+&2J!4#$*+(!@0(!B'$�'!VE!O#Ä$+"!:+$(Ä(*+"!eX(2+$@+2!X((0@'250(!-9;UJ!O674';+(7+59f!
%("N9>15(+25,67+!B+,,#()+(!*#$67)+8Q7$2E!/5+!%$)+;(5,,+!*5+,+$!B+,,#()+(!;+,2A25)2+(!*5+!
! ! ERGEBNISSE
`I!
K(('79+J!*',,!+,!,567!#9!K[L>109M+2525@+!X(75;520$+(!7'(*+&2E!O#;,2'("+(!4#$*+(!(#$!'&,!
V+52,2$#12#$+(!*+85(5+$2J!4+((!,5+!8Q$!*5+!%2cPWC!+5(+(!XcZD>_+$2!,045+!+5(+(!W5>_+$2!@0(!
1&+5(+$! GDD!ÜB! "+5)2+(! #(*! ,5+! +5(+!_',,+$&g,&5671+52! )$gr+$! ZDD!ÜB!;+,'r+(! ,045+! 57$+!
67+95,67+!O2$#12#$!952!VcoBO>!#(*!3BP>B+,,#()+(!;+,2A25)2!4+$*+(!10((2+(E!/5+!5*+(25>
85"5+$2+(!V+52,2$#12#$+(!,5(*!5(![';+&&+!YEF!'#8)+8Q7$2E!!
Tabelle 3.7: In vitro Experimente der identifizierten EtCRK2-Inhibitoren der zweiten Generation. !
G*'25*2'! =9)++.! B*4F=J!!/4TO!^kI_!
Vg+9#,-5.#*!
#%!@)++.'!^kI_!
!<+("595*'"0&>c'$;0(52$5&!
]H! ÉZDD!
!<+("595*'"0&>c'$;0(52$5&!
`C! ÉZDD!
!3'M720&'62'9! Z`! GCZ!
!3'M720&'62'9! `Z! GCZ!
!3'M720&'62'9! ]G! ÉZDD!
!3'M720&'62'9! ZY! ÉZDD!
!
?(M#;&5"5+$2+! %$)+;(5,,+! *+,! %("N9>Assays! @0(! B'$�'! VE! O#Ä$+"! :+$(Ä(*+"! eX(2+$@+2! X((0@'250(! -9;UJ!
O674';+(7+59fE!!
/5+!5(2+$+,,'(2+,2+!W&',,+!*+$!%2cPWC>X(75;520$+(!,5(*!*5+!<+("595*'"0&>c'$;0(52$5&+J!*'!,5+!
;5,&'()! 5(!*+$!V52+$'2#$!(#$!@0(!<'*'4+NJ!W'MM+!#(*!%&!U'4',7! x<K/K_%j!k!WKLL%J!
GHHHn!%V!UK_KOU et al.J!GHHHy!,045+!5(!+5(+9!L'2+(2!xKP<XO%P!k!KB<VKP/J!CDD`y!5(!
^#,'99+(7'()!952!*+$!'(25(+0M&',25,67+(!_5$1#()!*5+,+$!W&',,+!;+,67$5+;+(!4#$*+(E!K&&+!
'(*+$+(! O2$#12#$1&',,+(! ,5(*! ,670(! &A()+$+! ^+52! '&,! X(75;520$+(! *+$! c/W,! 0*+$! -&N60)+(>
,N(27',+(! e-OWf!;+1'((2! x<PKBOd3 et al.J! CDDGn!-PKj et al.J! GHHHn!VX? et al.J! CDDYyE!
/5+! :'95&5+(! *+$! c/W! #(*! *+$! -OW! ,5(*! +()! 952+5('(*+$! @+$4'(*2! xBK33X3- et al.J!
%
%
%% %
$
$& !&S!&
S
Q
%
$
$
%
$
%
!&S
!&S
Q
%
%
$$
$
!&S
Q
%
%
$$
$
$&
$&
%
%
%
$
!&S
Q
%
%$
$
$
!&S
!&S
ERGEBNISSE
`F!
CDDCyJ! *'7+$! 5,2! +,! ('7+&5+)+(*J! *',,! A7(&567+!O2$#12#$2NM+(! 5(! ;+5*+(!L$02+5(8'95&5+(! +5(+!
K125@52A2!"+5)+(E!!
Tabelle 3.8: In vitro Experimente der identifizierten Leitstrukturen der EtCRK2. !
/4TO!^kI_! =#^kI_!G*'25:*2'! =9)++.!
X+4A=J! B*4F=J! >&>=T! X+4A=J! B*4F=J!
Vg+9#,-:5.#*!#%!!
@)++.'!^kI_!
!
!!
d\5(*0&! DJCY!Å!DJGG! DJ``!Å!DJGH! CJID!Å!DJIF! DJDF!Å!DJDG! DJGI!Å!DJDY! ÉZDD!
!
!!
P70*'(5(>
d\5(*0&! C`!Å!GH! CY!Å!G]! C]!Å!I! GI!Å!F! G`!Å!H! ÉZDD!
!
!
3'M720&'62'9! GC!Å!I! Y`!Å!F! ÉCDD! C!Å!G! `!Å!C! ÉZDD!
!
!!
<+("59*5'"0&>
c'$;0(52$5&! ]G!Å!GY! `G!Å!CI! ÉCDD! ]G!Å!GY! FY!Å!CH! ÉZDD!
!
!!
<+("595*'"0&! YZ!Å!Y! GZ!Å!Y! ÉCDD! F!Å!C! F!Å!C! ÉZDD!
%,!10((2+(!@5+$!V+52,2$#12#$+(!@0(!X(75;520$+(!*+$!%2cPWC!;+,2A25)2!4+$*+(E!K&,!P+8+$+("!*5+(2+!+5(!d\5(*0&!
eX(*5$#;5(>Z>O#&80('2!x/KRX%O et al.J!CDDGyf!4+&67+,!+5(!X(75;520$!*+$!7#9'(+(!c/WC!5,2E!/+$!%("N9>Assay!
4#$*+!@0(!B'$�'!VE!O#Ä$+"!:+$(Ä(*+"! eX(2+$@+2! X((0@'250(!-9;UJ!O674';+(7+59f!*#$67)+8Q7$2E!K("'7&!*+$!
/#$678Q7$#()+(!59!%("N9+>Assayz!(!{!Y>]E!
!
PDJDS! G*'25*2':W5*#6#*<*+:".1#.-2%3.%!2%0!W%)98+.!0.'!>'$*.#%:V#3)%0.%:"#%02%3!^#$!67+95,67+(!dM2595+$#()!*+$!5*+(2585"5+$2+(!O2$#12#$+(!4#$*+(!Docking>%\M+$59+(2+!"#$!
K;&+52#()!@0(!O2$#12#$>K125@52A2,;+"5+7#()+(!,045+!"#$!R0$7+$,')+!*+$!<5(*+90*5!*#$67)+>
8Q7$2E!U5+$"#!4#$*+(!'&&+!*#$67!in vitro
! ! ERGEBNISSE
`H!
0(!"#$!Hinge>P+)50(E!:Q$!*5+!W&',,+!*+$!<+("595*'"0&>c'$;0(52$5&+!10((2+! p+*067!*+$!<5(>
*+90*#,!(5672!+5(*+#25)!@0$7+$)+,')2!4+$*+(E!!!
!
Abbildung 3.8: Analyse der Bindemodi der Benzimidazol-Carbonitrile.
U5+$!,5(*!*5+!"4+5!9g)&567+(!<5(*+90*5!*+$!<+("595*'"0&>c'$;0(52$5&+!*'$)+,2+&&2E!!%5(+!O#;,252#250(!'(!L0,525>
0(! PG! $+,#&25+$2! 59! 90&+1#&'$+(!Docking 5(! +5(+$! L0,+! *+$! <+("595*'"0&>c'$;0(52$5&+J! *5+! Q;+$! *+(! 352$5&>
O2561,2088!'(!*5+!Hinge>P+)50(!;5(*+(!e;&'#fE!O#;,252#250(+(!'(!L0,5250(!PY!+$"+#)+(!Docking>L0,+(J!;+5!*+(+(!
*5+!<+("595*'"0&>c'$;0(52$5&+! +5(+!_',,+$,2088;$Q61+!Q;+$! *+(!c'$;0(N&>O'#+$,2088! "#$!Hinge>P+)50(!;5&*+(!
e)$Q(fE!!
/5+!<5(*+90*5!*+$!<+("595*'"0&+!@'$55+$+(! 5(!Docking>K('&N,+(!';7A()5)!@09!O#;,252#25>
0(,9#,2+$!'(!L0,5250(!PGJ!PC!#(*!PY!e,5+7+!K;;5&*#()!YEFfE!%5(+!O#;,252#250(!'(!L0,5250(!PG!
#(*!PY! &5+8+$2+! 59!90&+1#&'$+(!Docking! +5(*+#25)+!<5(*+90*5E!/';+5! *5$5)5+$2! +5(! O#;,25>
2#+(2!'(!L0,5250(!PG!*+(!V5)'(*+(!,0J!*',,!+$!#(2+$!K#,;5&*#()!+5(+$!_',,+$,2088;$Q61+!Q;+$!
*+(!352$5&>O2561,2088! '(! *5+!Hinge>P+)50(! ;5(*+2E! %5(+! O#;,252#250(! '(! L0,5250(!PY! ;+45$12!
+5(+!P02'250(!*+,!V5)'(*+(!#9!GFDç!+(2&'()!*+$!70$5"0(2'&+(!K67,+!5(!*+$!L'M5+$+;+(+!e,5+7+!
[';+&&+!YEFfE!/'*#$67!;5(*+2!*+$!V5)'(*!Q;+$!*+(!c'$;0(N&>O'#+$,2088!#(2+$!K#,;5&*#()!+5(+$!
_',,+$,2088;$Q61+!'(!*5+!Hinge>P+)50(E!O#;,252#250(+(!'(!PC! 8Q7$2+(!"#!;+5*+(!9g)&567+(!
<5(*+90*5E!<+5!GDD!)+(+$5+$2+(!V5)'(*+(>L0,+(!952!O#;,252#+(2!'(!PC!4#$*+(!;+5*+!<5(*+>
%
%
$
%TS
TE
T)
ERGEBNISSE
ID!
90*5! '((A7+$(*!)&+5679Ar5)!@+$2+5&2! )+8#(*+(E!/5+!Scoring>_+$2+!#(2+$,675+*+(! ,567!(5672!
,5)(5851'(2E!%,!4'$!9522+&,!Docking!(5672!9g)&567!*5+!<+@0$"#)#()!+5(+,!<5(*+90*#,!('67>
"#@0&&"5+7+(E!^#$!)+('#+(!?(2+$,#67#()!*+,!<5(*+@+$7'&2+(,!4#$*+(!*+,7'&;!:$+5+>%(+$)5+>
O59#&'250(+(J! ,M+"5+&&!*5+!VX%>B+270*+! xKiRXO[!k!BKP%VX?OJ!CDDGyJ!+5()+,+2"2E!/5+,+!
5,2! 5(! *+$!V')+J! *5+! 8$+5+!<5(*#(),+(+$)5+! 8Q$!P+"+M20$>V5)'(*>W09M&+\+! "#!;+$+67(+(E! X(!
*+$!VX%>B+270*+!45$*!*+$!L$02+5(>V5)'(*>W09M&+\!@0&&,2A(*5)!8&+\5;+&!;+2$'672+2J!,045+!59!
<+$+567!*+$!V5)'(*><5(*+2',67+!,0&@'25,5+$2E!/'*#$67!5,2!+,!9g)&567J!%(2$0M5+;+52$A)+J!*5+!59!
Docking!('2#$)+9Ar!@+$('67&A,,5)2!4+$*+(J!"#!;+$Q61,56725)+(E!K#8)$#(*!*+$!R+$+5(8'67#(>
)+(J!*5+!*+$!VX%>B+270*+!"#)$#(*+!&5+)+(J!5,2!+,!(g25)J!*5+!%$)+;(5,,+!Q;+$!*5+!-+45672#(),>
8'120$+(! J! !#(*! !'(!+\M+$59+(2+&&!;+,25992+(!_+$2+(!'("#M',,+(E!!!
PDJDT! Q)9#0#.'2%3!0.'!V/B:I.*-$0.!2%0!".'.,-%2%3!0.'!K.E#,-*2%3+&)5*$'.%!^#$!R'&5*5+$#()!*+$!VX%>B+270*+!4#$*+!*5+!W$5,2'&&,2$#12#$!*+$!7#9'(+(!c/WC!eL/<>X/z!
GdXPf! 7+$'()+"0)+(E! /5+! V5)'(*+(*'2+(,A2"+! 4#$*+(! '#8! <',5,! @0(! @+$g88+(2&5672+(! <5(>
*#(),+(+$)5+*'2+(!+$,2+&&2E!U5+$"#!4#$*+(!"4+5!V5)'(**'2+(,A2"+!@+$4+(*+2E!/+$!+5(+!/'2+(>
,'2"! ,2'992!@0(!W0&;!et alE! x<PKBOd3 et al.J!CDDGn!-X<Od3 et al.J!CDDCn!WdV< et al.J!
CDDFy! *+$! '(*+$+!/'2+(,'2"! +(27A&2! V5)'(*+(! '#,! +5(+9! L'2+(2! xcUd3-!_%OV%j et al.J!
CDDYyE!U5+$;+5!4#$*+(!"#(A67,2!*5+!O2$#12#$+(!*+$!;+5*+(![+,2*'2+(,A2"+!9522+&,!Docking!5(!
*5+! W$5,2'&&,2$#12#$! +5()+M',,2J! #9! +5(+! K#,)'(),10(80$9'250(! +5(+,! 9g)&567+(! L$02+5(>
V5)'(*>W09M&+\+,! "#! +$7'&2+(E! O0407&! 8Q$! *+(!L'2+(2*'2+(,'2"! '&,! '#67! 8Q$! *+(!/'2+(,'2"!
@0(!W0&;!et alE!10((2+!+5(+!)#2+!W0$$+&'250(!"45,67+(!+\M+$59+(2+&&!;+,25992+(!<5(*#(),>
+(+$)5+(! #(*! 9522+&,! *+$! VX%>B+270*+! ;+$+67(+2+(! %(+$)5+(! +$"5+&2! 4+$*+(E! 3'67! W$+#">
R'&5*5+$#()!952!*+$!Leave-One-Out!B+270*+! x[dBX3K-K!k!=dP-%3O%3J!CDD]y!4#$*+!
+5(!iC!@0(!DJIG!8Q$!*+(!L'2+(2*'2+(,'2"!#(*!+5(!iC!@0(!DJIH!8Q$!*+(!/'2+(,'2"!@0(!W0&;!et alE!
+$$+5672E!/5+!*#$67,67(522&567+!K;4+567#()!@0(!*+(!+\M+$59+(2+&&!+$9522+&2+(!<5(*#(),+(+$>
)5+(!;+2$#)!DJ]G!16'&o90&! 8Q$!*+(!L'2+(2*'2+(,'2"!#(*!DJF`!16'&o90&! 8Q$!*+(!/'2+(,'2"!@0(!
W0&;!et alE!:Q$!*5+!VX%>B+270*+!45$*!+5(+!*#$67,67(522&567+!K;4+567#()!"45,67+(!+\M+$5>
9+(2+&&! +$9522+&2+9! #(*! ;+$+67(+2+9! ->_+$2! @0(! +24'! +5(+9! 16'&o90&! '()+(099+(!
x:dVdLL%!k!U?<<KP/J!CDD`yE!!!
! ! ERGEBNISSE
IG!
!
!!!
Abbildung 3.9: Scatter-Plots der Simulationsergebnisse von CDK-Inhibitoren mit der LIE-Methode.
Kf!VX%>O59#&'250(+(!8Q$!*+(!/'2+(,'2"!@0(!W0&;!et al.!x<PKBOd3 et al.J!CDDGn!-X<Od3 et al.J!CDDCn!WdV<
et al.J!CDDFyE!
<f!VX%>O59#&'250(+(!8Q$!*+(!L'2+(2*'2+(,'2"!xcUd3-!_%OV%j!et alEJ!CDDYyE!!!
^#$!K(M',,#()! *+$!-+45672#(),8'120$+(! *5+(2+! *',!U090&0)5+90*+&&! *+$! %2cPWCJ! ,045+!
;+1'((2+!c/W>X(75;520$+(!8Q$!*5+!in vitro!W5>_+$2+!952!*+$!%2cPWC!)+9+,,+(!4#$*+(E!/',!
VX%>B0*+&&!+$)';!+5(!iC!@0(!DJFI!5(!*+$!W$+#">R'&5*5+$#()!#(*!+5(+!*#$67,67(522&567+!K;>
4+567#()!@0(!DJYF!16'&o90&!@0(!*+(!+\M+$59+(2+&&!+$9522+&2+(!<5(*#(),+(+$)5+(E!%,!4#$*+(!
*5+!W0+885"5+(2+(! !{!DJFDJ! !{!DJYD!#(*! !{!>IJ`C!8Q$!*',!%2cPWC>VX%>B0*+&&!+$9522+&2E!!
<)W
<)W
<X
<X
<Y
<Y
<U
<U
<C
<C
<P
<))<)W<X<Y<U<C<P
!!
Abbildung 3.10: Scatter-Plots der LIE-Simulationen für die EtCRK2.
:Q$! *',! %2cPWC!U090&0)5+90*+&&!952! ;+1'((2+(!c/WC>X(75;520$+(J! 10((2+! +5(! M$A*5125@+,! VX%>B0*+&&!952!
+5(+9!iC!@0(!DJFI!+$,2+&&2!4+$*+(E!!
TE*Z*W[UXWE
<)S
<)E
<))
<)W
<X
<Y
<U
<C
<P
<)S<)E<))<)W<X<Y<U<C<P
TE*Z*W\U)EE
<)E
<))
<)W
<X
<Y
<U
<C
<)E<))<)W<X<Y<U<C
]E*Z*W\UX* ]
E*Z*W\U)*
]E*Z*W\YU*
1N487-F8.95"* ID-.?*36,5"^FA"B*
@_1*ID-.?*36,5"^FA"B*
0* /*
@_1*ID-.?*36,5"^FA"B*
@_1*ID-.?*36,5"^FA"B*
1N487-F8.95"* ID-.?*36,5"^FA"B* 1N487-F8.95"* ID-.?*36,5"^FA"B*
ERGEBNISSE
IC!
PDJDU! "#%0.($0#:Q$'-.'+)3.!0.'!".%1#(#0)1$9:4)';$%#*'#9.!!:Q$!*5+!<+("595*'"0&>c'$;0(52$5&+!952!O#;,252#250(!'(!L0,5250(!PC!4#$*+(!VX%>O59#&'250(+(!
*#$67)+8Q7$2E! U5+$8Q$! 4#$*+(! M$0! V5)'(*+(>L0,+! "4+5! VX%>O59#&'250(+(! +$,2+&&2E! /5+! <5(>
*#(),+(+$)5+(!4#$*+(!8Q$!p+*+!Docking>L0,+!e+5(9'&!Q;+$!*+(!c'$;0(N&>O'#+$,2088!Q;+$!+5(+!
_',,+$,2088;$Q61+!)+;#(*+(!'(!*5+!Hinge>P+)50(!#(*!+5(9'&!Q;+$!*+(!352$5&>O2561,2088!)+>
;#(*+(f! ;+$+67(+2E! /5+! ;+5*+(! <5(*#(),90*5! +5(+,! V5)'(*+(! 4#$*+(! '(7'(*! *+,! p+4+5&,!
;+,2+(!Docking-Scores!'#,)+4A7&2E!!
Tabelle 3.9: Bindemodi-Vorhersage der Benzimidazol-Carbonitirile mittels LIE-Methode.
!
G*'25*2'! "#%0.($02+! W%1)-9!A$,5#%3:>$+.%! Kl,)9!^5,)9i($9_!
d! ZZ! >IJHZ!!
! 3! ]Z! >FJ]G!
d! ZZ! >`JHY!!
!3! ]Z! >HJ]F!
d! CC! >]JZZ!!
!3! IF! >`JDI!
d! CZ! >]JYG!!
!3! IZ! >ZJFD!
d! HY! >YJYY!!
!3! I! >CJFI!
K#,!*+(!VX%>O59#&'250(+(! &+52+(!,567! &+5672+!?(2+$,675+*+! 5(!*+(!<5(*#(),+(+$)5+(!8Q$!*5+! p+4+5&5)+(!<5(*+>
90*5!';E!/5+,+!&5+)+(!p+*067!9+5,2!5(!*+$!-$gr+(0$*(#()!*+,!O2'(*'$*8+7&+$,!@0(!+5(+9!16'&o90&E!!!
%
%
$
!&S
%
!&S
%
%
%
%
$
$
%
%
%
$
$
$!&
S
%
%
%
$
$Q
$&
%
%
%
$
Q
!&S
! ! ERGEBNISSE
IY!
/5+!VX%>O59#&'250(+(!+$)';+(! &+5672+!?(2+$,675+*+! 5(!*+(!<5(*#(),+(+$)5+(!8Q$!*5+!;+5*+(!
<5(*+90*5! +5("+&(+$! V5)'(*+(E!/5+,+!?(2+$,675+*+! ,5(*! p+*067! "#9+5,2! 5(! *+$!-$gr+(0$*>
(#()!*+,!O2'(*'$*8+7&+$,!@0(!+5(+9!16'&o90&E!/$+5!:A&&+!;+4+)+(!,567!5(!+5(+$!-$gr+(0$*>
(#()!'#r+$7'&;!*+,!'()+(099+(+(!:+7&+$,J!p+*067!'#67!(5672!5(!+5(+$!-$gr+(0$*(#()J!*5+!+,!
+$&'#;2J! +5(+!d$5+(25+$#()! ,567+$! @0$7+$"#,')+(E!/5+!<5(*#()!Q;+$! *+(!352$5&>O2561,2088! "#$!
Hinge>P+)50(!,67+5(2!+(+$)+25,67!&+5672!;+@0$"#)2J!*+((067!,5(*!4'7$,67+5(&567!;+5*+!d$5+(>
25+$#()+(!9g)&567E!!
!
PDJDc! In silico!ZY*#(#.'2%3!0.'!".%1#(#0)1$9:4)';$%#*'#9.!
^#$! in silico! dM2595+$#()! *+$! <+("595*'"0&>c'$;0(52$5&+!4#$*+! +5(+! 109;5('20$5,67+! O#;>
,2'(";5;&5027+1!8Q$!*5+,+!O2$#12#$1&',,+!)+(+$5+$2E!U5+$"#!4#$*+!'(!*+(!L0,5250(+(!PG!;5,!PI!
,#;,252#5+$2! e,5+7+!K;,67(522! CEGC! ,045+!K;;5&*#()! CECfE!/5+!-+(+$5+$#()! *+$! 109;5('20$5>
,67+(!O#;,2'("*'2+(;'(1!4#$*+!9522+&,!*+$!O0824'$+!c'62@,J!45+!5(!W'M52+&!CEGC!;+,67$5+;+(J!
*#$67)+8Q7$2E!U5+$*#$67!4#$*+(!5(,)+,'92!GCEYZ`!O2$#12#$+(!952!*+$!-$#(*,2$#12#$!*+$!<+(>
"595*'"0&>c'$;0(52$5&+!)+(+$5+$2E!/5+!ON(27+25,5+$;'$1+52!4#$*+!8Q$!*5+! in silico!-+(+$5+$#()!
*+$!109;5('20$5,67+(!O#;,2'(";5;&5027+1!(5672!;+$Q61,56725)2E!/'7+$!7'(*+&2+!+,!,567!#9!+5(+!
$+5(! @5$2#+&&+! <5;&5027+1E! /5+! R+$;5(*#()+(! *5+,+$! @5$2#+&&+(! /'2+(;'(1! 4#$*+(! 5(! *',!
%2cPWC>U090&0)05+90*+&&! 9522+&,! Docking! +5()+M',,2E! %,! 4#$*+(! +(2,M$+67+(*! *+9! 5(!
K;,67(522!YECEC!;+,67$5+;+(+(!R0$)+7+(!p+4+5&,!GD!L0,+(!)+(+$5+$2!#(*!5(,)+,'92!GDD!)++5)>
(+2+! O#;,2'("+(! *#$67! C>O2#8+(><+4+$2#()J! X(2+$'1250(,85&2+$! ,045+! @5,#+&&+$! <+)#2'672#()!
'#,)+4A7&2E!K&,!4+52+$+,!K#,4'7&1$52+$5#9!4#$*+!*5+!Vg,&5671+52!*+$!O#;,2'("+(!l!;+$+67(+2!
952!R0&,#$8t!l!7+$'()+"0)+(E!3'67!Vg,&5671+52,'('&N,+(!@+$;&5+;+(!CD!R+$;5(*#()+(!5(!*+$!
85('&+(!K#,4'7&E!B52!*5+,+(!O2$#12#$+(!4#$*+(! 59!K(,67&#,,!<5(*#(),+(+$)5+(!9522+&,!*+$!
VX%>B+270*+!;+$+67(+2E
ERGEBNISSE
I]!
Tabelle 3.10: Optimierte Benzimidazol-Carbonitrile mit vorhergesagten Ki-Werten. !
ZY*#(#.'*.!G*'25*2'! K!^5,)9i($9_! =#!^%I_!
!>GGJHY! GJF!
!
>GDJH]! HJ`!
!
>GDJ`G! G`JH!
!
>GDJCF! CHJD!
!
>GDJDI! ]GJ`!
!
>GDJDD! ]IJY!
!
>HJZD! GDHJH!
!
>HJCD! GFGJZ!
!
>HJGC! CDHJY!
!>FJF]! YY]JD!
!
!
%
%
%
$%
%
&&
&
&
%
%
Q
%
$
&
$
$
%
%
%
% &&
&
&
&
&$
&
&
&
&
%
%
%
$
&
%%
&& &
&
%
%
Q
%
$
&
$
$
%
%
%
&
&
&
&$
&
& &
&
&
&
%
%
%
$
%
&
&
% &&
$
&
&
&&
%
%
Q
%
$
&
$
$
%
%
%
&
&
&
&
%
&&
%
%
%
$
&
$
&
% &&
#
#
#
%
%
%
$
%
&
&$
&%
&&
%
%
%
$
&
$
&
%$
$
&
&
%
%
%
$
%
&
&$
&%
&&
! ! ERGEBNISSE
IZ!
!!
Abbildung 3.11: Optimierte Benzimidazol-Carbonitril-Struktur.
/5+! 0M2595+$2+! O2$#12#$! ;5&*+2! "4+5!_',,+$,2088;$Q61+(! "#$!Hinge>P+)50(! '#,! ,045+! +5(+!_',,+$,2088;$Q61+!
"#9!-&#FF!#(*!'*$+,,5+$2!,0952!'#67!*5+!O+&+125@52A2,2',67+E!
!!PDP! /0.%*##.'2%3!6$%!B*IF=:!2%0!>&IF=:/%-#;#*$'.%!%5(!4+52+$+,!^5+&M$02+5(!"#$!%(24561&#()!(+#+$!K(25M'$',5251'!,5(*!*5+!BPW>7090&0)+(!W5>
(',+(!eBPW!{!MO15 Related Kinasesf!*+$!ApicomplexaE!/5+,+!L$02+5(+!)+7g$+(!+;+(8'&&,!"#!
*+$!:'95&5+!*+$!^N1&5(>';7A()5()+(!W5(',+(!#(*!4#$*+(!,0407&!8Q$!*5+!%2BPW!'&,!'#67!8Q$!
*5+!L8BPW!'&,!)++5)(+2+!^5+&M$02+5(+!@'&5*5+$2!e,5+7+!K;,67(522!GEZEZfE!%5(!^5+&!*5+,+$!K$;+52!
4'$!+,J!9522+&,!@5$2#+&&+(!Screenings!M02+(25+&&+!X(75;520$+(!*+$!L8BPW!#(*!%2BPW!"#!5*+(>
2585"5+$+(E!U5+$"#!4#$*+(!U090&0)5+90*+&&+!*+$!%2BPW!#(*!*+$!L8BPW!)+(+$5+$2!#(*!M0>
2+(25+&&+! X(75;520$+(!952!*+$!Docking>O0824'$+!LVK3[O!xWdP< et al.J!CDD`n!WdP< et al.J!
ERGEBNISSE
I`!
CDDHy! +5()+M',,2E! K&,! Screening>/'2+(;'(1! 4#$*+! *5+! <5(*5()/<! x444E;5(*5()*;E0$)y!
@+$4+(*+2E!/5+!<5(*5()/<!+(27A&2!;+1'((2+!O2$#12#$+(!,045+!K125@52A2,*'2+(!@0(!X(75;520$+(!
7#9'(+$!W5(',+(! xVX? et al.J!CDDIyJ!*5+!'#8!*5+,+!_+5,+! 8Q$!*+(!%5(,'2"!'&,!K(25M'$',5251'!
0M2595+$2!4+$*+(!1g((2+(E!!
!
PDPDH! X$($9$3#.($0.99#.'2%3!0.'!B*IF=!2%0!0.'!>&IF=!:Q$!*',!@5$2#+&&+!Screening! ('67!M02+(25+&&+(!%2BPW>!#(*!L8BPW>X(75;520$+(!4#$*+(!U0>
90&0)5+90*+&&+!*5+,+$!L$02+5(+!+$,2+&&2E!^#9!^+52M#(12!*+$!K$;+52+(!eO2'(*z!=#&5!CDDFf!,5(*!
5(! *+$! L$02+5(,2$#12#$*'2+(;'(1! L/<! x<%PBK3 et al.J! CDDYy! 8Q(8! W$5,2'&&,2$#12#$+(! @0(!
c/W>A7(&567+(! L$02+5(+(! *+$!Apicomplexa! e+5(+! @0(!Cryptosporidium parvum! ,045+! @5+$!
@0(! P. falciparumf! @+$"+567(+2J! *5+! '&,! M02+(25+&&+! Template! 8Q$! *5+! B0*+&&5+$#()! *5+(+(!
1g((2+(E!/5+,+!L$02+5(+!,5(*!7090&0)!"#!U,c/WCE!/+,!_+52+$+(!4'$+(!GHG!W$5,2'&&,2$#12#>
$+(!*+$!U,c/WC!#(*!+5(+!W$5,2'&&,2$#12#$!*+$!U,c/WI!5(!*+$!L/<!';)+&+)2E!O0952!,2'(*+(!
5(,)+,'92! GHI! [+9M&'2,2$#12#$+(! 8Q$! *5+! U090&0)5+90*+&&5+$#()! *+$! %2BPW! #(*! *+$!
L8BPW!"#$!R+$8Q)#()E!=+*067!,0&&2+!8Q$!*5+!B0*+&&5+$#()!+5(+!W$5,2'&&,2$#12#$!'&,![+9M&'2!
)+4A7&2!4+$*+(J!*5+!+5(+!9g)&567,2!707+!O+b#+("5*+(252A2!"#!*+$!Target>L$02+5(,+b#+("!'#8>
4+5,2E!X(![';+&&+!YEGG!,5(*!9g)&567+![+9M&'2,2$#12#$+(!952!*+(!p+4+5&5)+(!O+b#+("5*+(252A2+(!
"#!*+$!%2BPW!#(*!*+$!L8BPW!'()+)+;+(E!!!
/5+!5(!*+$!L/<!';)+&+)2+(!U,c/WC!W$5,2'&&,2$#12#$+(!&5+)+(!"#9![+5&!59!'125@+(!W09M&+\!
'&,!U+2+$0*59+$J!*E!7E!)+;#(*+(!952!+5(+9!^N1&5(!@0$J!#(*!,5(*!2+5&4+5,+J!9522+&,!L70,M70$N>
&5+$#()!'(![7$G`D!59!K125@5+$#(),>LoopJ!@0&&,2A(*5)!'125@5+$2E!:Q$!*',!,2$#12#$;',5+$2+!_5$1>
,2088*+,5)(!,5(*! 5(,;+,0(*+$+!O2$#12#$+(! 5(2+$+,,'(2J!*5+!952!+5(+9!X(75;520$!60>1$5,2'&&5,5+$2!
,5(*E! /5+,+! O2$#12#$+(! 4+5,+(! +5(+! L$02+5(10(80$9'250(! '#8J! *5+! *#$67! *5+! X(2+$'1250(! 952!
*+9! V5)'(*+(! "#,2'(*+! 1099+(! eInduced Fitf! x%3/Xcd[[ et al.J! GHHHn! 3d<V% et al.J!
CDD]yE!/5+,+!O2$#12#$9+$19'&+!+$9g)&567+(!*5+!)+"5+&2+!%(24561&#()!(+#+$! X(75;520$+(!952!
+5(+$!8Q$!*5+!L$02+5(;5(*+2',67+!0M2595+$2+(!O2$#12#$J!#(*!+5)(+(!,567!*'7+$!;+,0(*+$,!)#2!'&,!
[+9M&'2,2$#12#$! 8Q$! *5+! U090&0)5+90*+&&5+$#()! ,045+! '#67! 8Q$! *5+! K('&N,+! @0(! L$02+5(>
V5)'(*>_+67,+&45$1#()+(E!/'$Q;+$!75('#,! ,0&&2+! +5(+
! ! ERGEBNISSE
II!
x<P%X[%3V%cU3%P et al.J!CDDZyn!x3XOOX3W et al.J!CDDCyE!K#r+$*+9!"+5)2!*5+,+!W$5,2'&&>
,2$#12#$! +5(+! )++5)(+2+! W0(80$9'250(! *+,! K95(0,A#$+$+,2+,! VN,YY! eU,c/WC>
3#99+$5+$#()fJ!*5+!+5(+!50(5,67+!_+67,+&45$1#()!952!M02+(25+&&+(!X(75;520$+(!+$&'#;2E!K(>
$+567+$#(),+\M+$59+(2+!;+1'((2+$!V5)'(*+(!9522+&,!Docking!"+5)+(!)#2+!%$)+;(5,,+!952!*5+>
,+$!W$5,2'&&,2$#12#$!x<%j%PJ!CDDIyE!!!
Tabelle 3.11: Mögliche Templatstrukturen zur Homologiemodellierung der EtMRK und der PfMRK
!
B*IF=! >&IF=!
Z'3)%#+(2+!/0.%*#*<*!
^j_!
d-%9#,-5.#*!
^j_!
/0.%*#*<*!
^j_!
d-%9#,-5.#*!
^j_!
>A":
/A!
W2&9g+2%3!
^m_!
cMc/WC! YZ! ZZ! YH! Z`! CiWP! CJ`!
L8LWZ! Y`! ZG! ]C! ZY! GRDd! GJH!
U,c/WC! YF! ZH! YH! ZC! GdXP! GJH!
U,c/WI! YF! `C! ]D! ZC! G?KC! YJD!
^#$!-+(+$5+$#()! *+$!U090&0)5+90*+&&+! 8Q$! *5+! %2BPW! #(*! *5+! L8BPW!4#$*+! *5+! 7#9'(+! c/WC! O2$#12#$!
eL/<>K66E>3$Ez! GdXPfJ! 2$02"!*+$! +24',!)+$5()+$+(!O+b#+("A7(&5671+52! 59!R+$)&+567! "#$! 7#9'(+(!c/WI!@+$>
4+(*+2J!*'!*5+,+!O2$#12#$!+5(+!,+7$!)#2+!K#8&g,#()!;+,52"2E!!
^#! <+)5((! *+$!U090&0)5+90*+&&5+$#()!4#$*+(! *5+! O+b#+("+(! @0(! GdXP! #(*! *+$! %2BPW!
,045+!*+$!L8BPW!'&5)(5+$2!#(*!*'((!p+4+5&,!"+7(!B0*+&&+!952!*+$!O0824'$+!Bd%!)+(+$5+$2E!
/5+!B0*+&&+!4#$*+(!'(,67&5+r+(*!'(7'(*!+5(+$!L$02+5(>-+09+2$5+>K('&N,+! 5(!Bd%!;+4+$>
2+2E!/';+5!4#$*+!"#(A67,2!+5(!P'9'67'(*$'(>L&02!8Q$!;+5*+!B0*+&&+!+$,2+&&2!e,5+7+!K;;5&*#()!
YEGCfE!X(!*5+,+9!L&02!4+$*+(!*5+! >!#(*! >!_5(1+&!*+$!K95(0,A#$+(!)+)+(+5('(*+$!'#8)+>
2$')+(E!/5+,+$!L&02!+$9g)&5672!+5(+!+5(8'67+!)$'M75,67+!K#,,')+!Q;+$!8'@0$5,5+$2+!#(*!+$&'#;>
2+!<+$+567+! +5("+&(+$!K95(0,A#$+(E!K95(0,A#$+(J! *5+! ,567! 5(! (5672>8'@0$5,5+$2+(!<+$+567+(!
;+85(*+(J!,0&&2+(!)+('#+$!Q;+$M$Q82!4+$*+(E!_A7$+(*!;+59!%2BPW>B0*+&&!'&&+!K95(0,A#$+(!
)Q(,25)+! [0$,50(,45(1+&! #(*! ,0952! (5+*$5)+! %(+$)5+(! ;+,52"+(J! 85(*+(! ,567! ;+59! L8BPW>
B0*+&&! @5+$! K95(0,A#$+(!952! #()Q(,25)+(! [0$,50(,45(1+&(! e,5+7+! K;;5&*#()! YEGCfE! /5+,+!
&5+)+(!p+*067!'#r+$7'&;!*+$!L$02+5(;5(*+2',67+!#(*!4#$*+(!*'7+$!(5672!4+52+$!;+$Q61,56725)2E!
ERGEBNISSE
IF!
!
!
Abbildung 3.12: Ramachandran-Plots der Homologiemodelle der EtMRK und der PfMRK.
<+$+567+! 8'@0$5,5+$2+$! [0$,50(,45(1+&! ,5(*! )$Q(! #9$'(*+2E!-+&;! #9$'(*+2! ,5(*!<+$+567+J! *+$+(!K95(0,A#$+(!
+$&'#;2+
! ! ERGEBNISSE
IH!
!
Abbildung 3.13: Bindungsmodus eines bekannten PfMRK-Inhibitors.
/+$!;+1'((2+!d\5(*0&+>X(75;520$!x_dd/KP/!et alEJ!CDDYy!;5(*+2!Q;+$!2NM5,67+!_',,+$,2088;$Q61+(;5(*#()+(!
"#$!Hinge>P+)50(E!!
!
PDPDJ! B6)92#.'2%3!0.'!Docking:G$&*E)'.!>VW?MG!
/5+! Docking>O0824'$+! LVK3[O! +$9g)&5672! +,J! +5(+! @5$2#+&&+! Screening>/'2+(;'(1! ,+7$!
,67(+&&!('67!M02+(25+&&+(!V5)'(*+(!"#!*#$67,#67+(!#(*!4#$*+!*'7+$!"#9!@5$2#+&&+(!Screening!
('67! X(75;520$+(! *+$! %2BPW! ,045+! L8BPW! +5()+,+2"2! xWdP< et al.J! CDD`n! WdP< et al.J!
CDDHyE!/'!;5,&'()!(#$!4+(5)+!@5$2#+&&+!Screenings!952!*5+,+$!Docking>O0824'$+!*#$67)+8Q7$2!
4#$*+(J!9#,,2+!*5+!O0824'$+!"#(A67,2!75($+567+(*!+@'+$2!4+$*+(E!O5+!4#$*+!*'7+$!5(!+5(5>
)+(!Docking>%\M+$59+(2+(!952! *+$!Docking>O0824'$+!-dV/! x=d3%O et al.J! GHHZy! @+$)&5>
67+(E!
!
ERGEBNISSE
FD!
YEYECEG* P+>Docking!%\M+$59+(2+!
^#$!%@'+$#()!*+$!Docking>O0824'$+!LVK3[O!4#$*+(!"#(A67,2!P+>Docking!%\M+$59+(2+!
*#$67)+8Q7$2!#(*!952!*+$!O2'(*'$*>Dockingsoftware!-dV/!@+$)&567+(E!:Q$!+5(!P+>Docking!
%\M+$59+(2!;+(g25)2!9'(!)++5)(+2+!W$5,2'&&,2$#12#$+(!952!60>1$5,2'&&5,5+$2+(!V5)'(*+(E!U5+$"#!
4#$*+(! GZ! W$5,2'&&,2$#12#$+(! *+$! 7#9'(+(! c/WC! @+$4+(*+2J! *5+! +5(+! K#8&g,#()! 1&+5(+$!!
CJZ! h! ;+,52"+(J! 952! +5(+9! X(75;520$! 60>1$5,2'&&5,5+$2! ,5(*! #(*! *+$+(! <5(*+2',67+(>
K95(0,A#$+(! @0&&,2A(*5)! 15$,2'&&0)$'M75,67! '#8)+&g,2! ,5(*E! :Q$! *',!P+>Docking!4#$*+(! *5+!
X(75;520$+(! "#(A67,2! '#,! *+$! L$02+5(;5(*+2',67+! +(28+$(2! #(*! (+#+! O2'$210(80$9'250(+(! ;+>
$+67(+2!xOK/d_OWXJ!GHHIn![%[WdJ!CDDZyE!:Q$!*+(!R+$)&+567!"45,67+(!?$,M$#(),10(80$>
9'250(!*+$!V5)'(*+(!#(*!W0(80$9'250(!*+$!V5)'(*+(!('67!*+9!DockingJ!4#$*+(!root mean
square deviation! ePBO/f!_+$2+! *+$!O674+$'209+!*+$!V5)'(*+(! ;+$+67(+2! #(*! *5+(2+(! '&,!
B'r!8Q$!*5+!i#'&52A2!*+$!L0,+(>-+(+$5+$#()!*+,!Docking-K&)0$5279#,E!=+!1&+5(+$!*+$!$+,#&25+>
$+(*+!PBO/>_+$2J!*+,20!)+('#+$!$+M$0*#"5+$2!*5+!Docking>O0824'$+!*5+!V5)'(*+(>L0,+E!K&,!
+5(!'#,$+567+(*!)+('#+,!Docking!4#$*+!+5(!PBO/>_+$2!@0(!9'\59'&!C!h!'()+(099+(E!:Q$!
*5+,+!%\M+$59+(2+!4#$*+(!*5+!%5(,2+&&#()+(!*+$!LVK3[O>!#(*!*+$!-dV/>O0824'$+!,0!)+>
4A7&2J!*',,! ,5+!+5(+!9g)&567,2! ,67(+&&+!<+$+67(#()!*+$! +5("+&(+(!V5)'(*+(!+$&'#;+(! eSree-
ning!B0*#,fE!:Q$!p+*+(!V5)'(*+(!4#$*+(!CZ!@+$,675+*+(+!L0,+(!)+(+$5+$2!#(*!59!K(,67&#,,!
*'$'(!B522+&4+$2+! *+$! ;+$+67(+2+(! PBO/>_+$2+! )+;5&*+2! e,5+7+! K;;5&*#()! YEG]fE! /5+! %$>
80&),4'7$,67+5(&5671+52J! +5(+! V5)'(*+(>L0,+! 952! +5(+9! /#$67,67(522,>PBO/! @0(! 1&+5(+$!!
C!hJ!@+$)&567+(!952!*+$!W0(80$9'250(!*+$!W$5,2'&&,2$#12#$J!"#!)+(+$5+$+(J!&')!;+5!CIq!8Q$!*5+!
LVK3[O>O0824'$+!,045+!;+5!ZYq!8Q$!*5+!-dV/>O0824'$+E!/5+!$+&'25@!707+!O2'(*'$*';4+5>
67#()! *+$! /#$67,67(522,>PBO/>_+$2+! *+$! +5("+&(+(! L$02+5(>V5)'(*! W09M&+\+! &5+)2! #(2+$!
'(*+$+9!*'$5(!;+)$Q(*+2J!*',,!5(!+5(5)+(!%\M+$59+(2+(!,+7$!#(2+$,675+*&567+!L0,+(!*+,,+&;+(!
V5)'(*+(!@0$7+$)+,')2!4#$*+(J!4+&67+!*+9+(2,M$+67+(*!707+!PBO/>_+$2+!'#845+,+(E!%5>
(5)+!X(75;520$+(!"+5)2+(!"#*+9!Docking>L0,+(!*5+!#9!GFDç!$025+$2!@0$&')+(E!?(2+$,675+*&567+!
<5(*+90*5!5(!Docking>%\M+$59+(2+(J!85(*+(!,567!8Q$!c/W>X(75;520$+(!7A#85)J!*'!,5+!59!Do-
cking!952!*+$!Hinge>P+)50(!Q;+$!@+$,675+*+(+!_',,+$,2088;$Q61+(>B#,2+$!5(!X(2+$'1250(!2$+>
2+(!1g((+(!x_? et al.J!CDDYy!#(*!;+5!+5(+$!/$+7#()!#9!GFDç!,0)'$!*',!2NM5,67+!X(2+$'1250(,>
B#,2+$!952!*+$!Hinge>P+)50(!;+5;+7'&2+(E!!
! ! ERGEBNISSE
FG!
!
Abbildung 3.14: Durchschnitts-RMSD-Werte der Liganden-Posen nach einem Re-Docking mit der Software
PLANTS und GOLD.
/5+!<+5*+(!-$'851+(!@+$'(,67'#&567+(!*+#2&567J!*',,!*5+!Docking>O0824'$+!-dV/!7A#85)+$!*5+!;50'125@+!W0(>
80$9'250(!$+M$0*#"5+$+(!10((2+E!!!
X(!*+9!<'&1+(*5')$'99!eK;;5&*#()!YEGZf! $+M$A,+(25+$+(!*5+!<'&1+(!*5+!K("'7&!*+$! $56725)!
)+(+$5+$2+(!Docking>L0,+(E!/5+!-dV/>O0824'$+!4'$! 5(!``q!'&&+$!)+(+$5+$2+(!L0,+(! 5(!*+$!
V')+J!*5+!L$02+5(>V5)'(*>W09M&+\+!+$80&)$+567!"#! $+M$0*#"5+$+(E!%5(+!A7(&567!)#2+!Perfor-
W[W
)[W
E[W
S[W
2[W
P[W
C[W
U[W
Y[W
X[W
)0])
)')Y
)1)`
)I=Y
)&W)
)aQ`
)a`b
)(1X
)$_]
)$_T
)$_c
)bd_
)TUY
)cTe
)`W$
K<"b!cde
b"5.9:
W[W
)[W
E[W
S[W
2[W
P[W
C[W
U[W
Y[W
X[W
)0])
)')Y
)1)`
)I=Y
)&W)
)aQ`
)a`b
)(1X
)$_]
)$_T
)$_c
)bd_
)TUY
)cTe
)`W$
K<"b!cde
IA"?
L/<>Accession-3#99+$!
L/<>Accession-3#99+$!
ERGEBNISSE
FC!
mance! 4#$*+! '#67! @0(! W+&&+(;+$)+$! et al.! ;+,67$5+;+(! xW%VV%3<%P-%P et al.J! CDD]yE!
LVK3[O!+$"5+&2+!(#$!5(!YYq!'&&+$!)+(+$5+$+(!L0,+(!+5(+(!/#$67,67(522,>PBO/!@0(!1&+5(+$!C!
hJ! @+$)&567+(!952! *+$!W0(80$9'250(! *+$!W$5,2'&&,2$#12#$E! :Q$! +5(! @5$2#+&&+,!Screening! 5,2! +,!
(5672!(024+(*5)J!*',,!'&&+!)+(+$5+$2+(!V5)'(*+(>L0,+(!+5(+,!+5("+&(+(!V5)'(*+(!*+$!;50'125>
@+(! W0(80$9'250(! +(2,M$+67+(E!_56725)! 5,2J! *',,! *5+! @+$4+(*+2+! <+4+$2#(),8#(1250(! "45>
,67+(!S$56725)T!#(*!S8'&,67T!)+(+$5+$2+(!Docking>L0,+(!*5,1$595(5+$+(!1'((!xcPdOO et al.J!
CDDHyE!^#*+9!4#$*+(!Docking>L'$'9+2+$!@+$4+(*+2J! *5+! +5(+! ,67(+&&+!<+$+67(#()!*+$!V5>
)'(*+(>L0,+(!+$&'#;+(E!:Q$!*5+!W$5,2'&&,2$#12#$!GUDG!85(*+2!LVK3[O!5(!CZ!)+(+$5+$2+(!Do-
cking>L0,+(!1+5(+!10$$+12+!L0,+E!/5+!O0824'$+!-dV/!4'$!75()+)+(!(5672! 5(!*+$!V')+!+5(+!
10$$+12+!V5)'(*+(>L0,+!*+$!W$5,2'&&,2$#12#$!GdXP!"#!)+(+$5+$+(E!^#!+5(+9!A7(&567+(!%$)+;>
(5,!1'9!<+N+$!#(2+$!R+$4+(*#()!*+$!O0824'$+!K#20/061!YED!x<%j%PJ!CDDIyE!/5+,!1g((2+!
'#8! :+7&+$! 5(! *+(! L$02+5(,2$#12#$+(! "#$Q61"#8Q7$+(! ,+5(! xOd[PX::%P! k! /PKB<?P-J!
CDDZyE!!
!
Abbildung 3.15: Anzahl der richtig reproduzierten Liganden-Posen nach Re-Docking Experimenten mit der
Software PLANTS und GOLD.
/5+!Docking>O0824'$+!-dV/!85(*+2!59!-+)+(,'2"!"#!LVK3[O!7A#85)+$!10$$+12+!V5)'(*+(>L0,+(J!*5+!*+$!W$5,>
2'&&,2$#12#$!+(2,M$+67+(E!:Q$!,+67,!*+$!GZ!W$5,2'&&,2$#12#$+(!4#$*+!,2+2,!*5+!10$$+12+!V5)'(*+(>L0,+!+$9522+&2E!!!
3+;+(! *+$! -+(+$5+$#()! +5(+$! 10$$+12+(! V5)'(*+(>L0,+J! 4#$*+(! *5+! ;+5*+(! Docking>
K&)0$5279+(!@0(!-dV/!#(*!LVK3[O!'#67!5(!<+"#)!'#8!57$+!<+4+$2#(),8#(1250(+(!)+2+,>
2+2E!U5+$;+5!,0&&2+!Q;+$M$Q82!4+$*+(J!0;!*5+!-Q2+!*+$!+$9522+&2+(!V5)'(*+(>L0,+(!'#67!952!*+$!
5(2+$(+(!<+4+$2#(),8#(1250(! *+$!Docking>O0824'$+! 10$$+&5+$2E! /'"#!4#$*+! *',! 85('&+!Ran-
W
P
)W
)P
EW
EP
SW
)0])
)')Y
)1)`
)I=Y
)&W)
)aQ`
)a`b
)(1X
)$_]
)$_T
)$_c
)bd_
)TUY
)cTe
)`W$
:/*1#!bGb"5.9: IA"?
L/<-Accession>3#99+$!
! ! ERGEBNISSE
FY!
king!@5'!PBO/>_+$2!#(*!@5'!Scoring>_+$2!952+5('(*+$!@+$)&567+(!#(*!+5(+!OM+'$9'(!W0$>
$+&'250(!x_XOOV%PJ!GHDZy!*#$67)+8Q7$2E!<+5!*5+,+$!K$2!@0(!W0$$+&'250(,'('&N,+!45$*!#(2+$>
,#672J! 5(45+4+52!"4+5!R'$5';&+(!e"E!<E!Scoring>!#(*!PBO/>_+$2f! 5(!<+"5+7#()!"#+5('(*+$!
,2+7+(E!/+$!75+$;+5!;+$+67(+2+!OM+'$9'(!W0$$+&'250(,10+885"5+(2!P,!1'((!"45,67+(!>G!#(*!G!
@'$5+$+(!e>G!{!5(@+$,+!W0$$+&'250(J!D!{!1+5(+!W0$$+&'250(J!G!{!W0$$+&'250(fE!P,!,0&&2+!9g)&567,2!
('7+!+5(,!&5+)+(E!/5+,!4Q$*+!;+*+#2+(J!*',,!$56725)+!V5)'(*+(>L0,+(!+5(+(!;+,,+$+(!Docking-
Score!+$7'&2+(!'&,!,0&67+!952!(5672!10$$+12+(!L0,+(E!<+5!+5(+9!(+)'25@+(!P,!75()+)+(!4Q$*+(!
,67&+672+$+!V5)'(*+(>L0,+(!;+,,+$!;+4+$2+2!4+$*+(!'&,!10$$+12+!L0,+(E!/5+!K;;5&*#()!YEG`!
"+5)2J!*',,!*5+! 5(2+$(+!<+4+$2#(),8#(1250(!@0(!LVK3[O!)#2!)++5)(+2! 5,2J!#9!+5(+!)#2+!V5>
)'(*+(>L0,+!"#!5*+(2585"5+$+(E!:Q$!,5+;+(!*+$!GZ!P+>Scoring>%\M+$59+(2+!+$$+67(+2+!,567!+5(!
W0$$+&'250(,10+885"5+(2!P,!@0(!Q;+$!DJZE!/+$!/#$67,67(522,>P,!Q;+$!'&&+!%\M+$59+(2+! &')!8Q$!
LVK3[O! ;+5! DJYCE! /5+!Docking>O0824'$+!-dV/! ,67(522! 75+$!952! +5(+9!/#$67,67(522,>! P,!
@0(!DJDF!*+#2&567!,67&+672+$!';!#(*!"+5)2+!59!/#$67,67(522!1+5(+!W0$$+&'250(!"45,67+(!*+9!
Docking-Score! #(*!*+9!PBO/>_+$2E! X(! +&8! @0(!GZ!P+>Docking>%\M+$59+(2+(!4'$!P,!@0(!
LVK3[O!7g7+$!'&,!;+5!-dV/!#(*!(#$! 5(!"4+5!P+>Docking>%\M+$59+(2+(! e8Q$!*5+!W$5,2'&&>
,2$#12#$+(!G=OR!#(*!GL}Xf!,67(522!-dV/!;+,,+$!';E!
!
Abbildung 3.16: Vergleich zwischen dem RMSD-Wert und der Anreicherung von aktiven Substanzen.
/5+!-$'851!"+5)2J!*',,!*5+!Scoring>:#(1250(!@0(!LVK3[O!7A#85)+$!+5(+!10$$+12+!Docking>L0,+!5*+(2585"5+$2E!/5+!
10$$+12+(!Docking>L0,+(!e*E!7E!L0,+(!952!+5(+9!PBO/!@0(!1&+5(+$!C!hf!;+1099+(!+5(+(!;+,,+$+(!Docking-
ScoreE!!
!
<)[W
<W[Y
<W[C
<W[2
<W[E
W[W
W[E
W[2
W[C
W[Y
)[W
)0])
)')Y
)1)`
)I=Y
)&W)
)aQ`
)a`b
)(1X
)$_]
)$_T
)$_c
)bd_
)TUY
)cTe
)`W$
K( b"5.9: IA"?
L/<-Accession>3#99+$
ERGEBNISSE
F]!
%(2,67+5*+(*!8Q$!+5(!@5$2#+&&+, Screening!5,2J!*',,!*5+!V5)'(*+(>L0,+!952!*+9!;+,2+(!Scoring>
_+$2J!+5(+(!PBO/>_+$2!@0(!1&+5(+$!C!h!;+,52"2!#9!L0,+(J!*5+!(5672!*+$!;50'125@+(!W0(80$>
9'250(! +(2,M$+67+(J! "#! +(28+$(+(E! /5+! -dV/>O0824'$+! 4'$! 8Q$! IYq! *+$! L$02+5(>V5)'(*>
W09M&+\+!)++5)(+2J!+5(+!A7(&567+!;50'125@+!W0(80$9'250(!"#!,+&+125+$+(E!LVK3[O!+$"5+&2+!5(!
HYq!*+$!:A&&+!+5(+!O+&+1250(!*+$!;50'125@+(!W0(80$9'250(!e,5+7+![';+&&+!YEGCfE!
Tabelle 3.12: RMSD-Werte der Liganden-Posen mit dem besten GOLD-Score sowie PLANTS-Score nach einem
Re-Docking in die Proteinbindetasche. !
-dV/>Score! LVK3[O>Score!L/<>X/!
PBO/! PBO/!
GKiG! GJC! DJH!
G/GF! IJG|! GJC!
G%GR! DJ]! DJ`!
G-^F! GJD! DJF!
GUDG! CJG|! ]J]|!
G=OR! GJZ! GJG!
G=RL! DJ`! DJZ!
GW%H! GJZ! GJZ!
GdXi! DJI! DJI!
GdXP! FJY|! GJI!
GdX?! DJF! DJF!
GL}X! DJY! DJI!
GPIF! GJ]! GJZ!
G?P_! CJH|! CJD!
GRDd! DJI! DJZ!
K()+)+;+(! 5,2! *+$!PBO/>_+$2! *+$! V5)'(*+(>L0,+! +5(+,!P+8+$+(">V5)'(*+(J! *5+! @0(! *+(! +5("+&(+(!Scoring>
:#(1250(+(!'9!;+,2+(!;+4+$2+2!4#$*+E!
! ! ERGEBNISSE
FZ!
YEYECEC* K($+567+$#(),+\M+$59+(2+!
^#$!4+52+$+(!%@'+$#()! *+$!Software!-dV/!#(*!LVK3[O!4#$*+(!K($+567+$#(),+\M+$5>
9+(2+! @0(! ;+1'((2+(! X(75;520$+(! *+$! 7#9'(+(! c/WC! *#$67)+8Q7$2E! L$5("5M5+&&! ,0&&2+! +5(+!
Docking>O0824'$+!59!in silico!Screening!)++5)(+2!,+5(J!"45,67+(!V5)'(*+(!+5(+,!L$02+5(,!#(*!
,0&67+(J! *5+! 4'7$,67+5(&567! (5672! ;5(*+(! "#! #(2+$,67+5*+(E! /+,7'&;! 4#$*+(! Docking>
%\M+$59+(2+!952!+5(+9!,M+"5+&&!8Q$!,0&67+!:$')+,2+&&#()+(!+(24561+&2+(![+,2*'2+(,'2"!e,5+7+!
CECn!/?/>[+,2*'2+(,'2"J!xU?K3- et al.J!CDD`yf!952!*+9!c/WC>L$02+5(!*#$67)+8Q7$2E!/5+,+$!
/'2+(,'2"!+(27A&2!IC!c/W>X(75;520$+(!#(*!5(,)+,'92!CEDI]!5('125@+!R+$;5(*#()+(E!/5+!X('1>
25@52A2!8Q$!*5+,+!L$02+5(1&',,+!4#$*+!in silico!@0$7+$)+,')2!#(*!4#$*+!(5672!in vitro!;+,2A25)2E!
K&,! L$02+5(,2$#12#$! 8Q$! *',! Docking! *5+(2+! 45+*+$#9! +5(+! O2$#12#$! *+$! humanen! c/WC!
eL/<>K66E>3$Ez!GdXPfE!/5+!%5(,2+&&#()+(!*+$!L'$'9+2+$!@0(!LVK3[O!#(*!-dV/!+(2,M$'>
67+(!*+(+(!+5(+,!@5$2#+&&+(!Screening!SetupsE!!!
E
Ef=
Ef?
Ef;
Ef@
EfA
EfF
EfB
EfC
EfD
=
E Ef= Ef? Ef; Ef@ EfA EfF EfB EfC EfD =
K$#)!0).!5$'(VWMG/(6#6\)1
K$#)!0).!K6VW#62MG/(6#6\)1
7/'0G'$1#(
!
Abbildung 3.17: ROC-Kurve
/5+!;+5*+(!)+(+$5+$2+(!Pdc>W#$@+(!8Q$!*',!LVK3[O>!#(*!-dV/>Docking!"+5)+(J!*',,!*5+!Scoring>:#(1250(!
LVK3[O>Score! 59! -+)+(,'2"! "#! -dV/>Score! ;+,,+$! 5(! *+$! V')+! 4'$J! "45,67+(! SP56725)>L0,525@+(T! e*E7E!
c/W>X(75;520$+(f!@0(!S:'&,67>L0,525@+(T!eDecoysf!"#!*5,1$595(5+$+(E!!!
^#$!K#,4+$2#()! *+,!K($+567+$#(),>%\M+$59+(2+,! *5+(2+! *5+!Pdc>W#$@+! ePdc!{!Receiver
Operating CharacteristicfE!X(!*5+,+$!45$*!*5+!P'2+!*+$!S:'&,67>L0,525@+(T!e5('125@+!V5)'(*+(f!
ERGEBNISSE
F`!
952! 707+9!Docking-Score! "#$! P'2+! *+$! SP56725)>L0,525@+(T! e'125@+! V5)'(*+(f! '#8)+2$')+(E!
K;;5&*#()!YEGI!"+5)2!*+(!R+$)&+567!"45,67+(!*+(!;+5*+(!Pdc>W#$@+(!8Q$!*',!LVK3[O>!#(*!
*',! -dV/>DockingE! %5(+! )#2+! K($+567+$#()! @0(! '125@+(! O#;,2'("+(! 10((2+! 952! *+$!
LVK3[O>O0824'$+!+$"5+&2!4+$*+(E!/5+!-dV/>O0824'$+! ,67(522!;+5!*+(!*#$67)+8Q7$2+(!Do-
cking>%\M+$59+(2+(! ,67&+672+$! ';E!%5(!B'r! 8Q$! *5+!-Q2+! *+,![+,2,! 5,2! *5+!:&A67+!#(2+$! *+$!
Pdc>W#$@+! eK?c!{!Area Under CurvefE!/5+!:&A67+!1'((!_+$2+!"45,67+(!D!#(*!G!'((+7>
9+(J!40;+5! +5(! 7g7+$+$!_+$2! +5(+! 7g7+$+!-Q2+! '("+5)2E!B52! LVK3[O! 10((2+! +5(! ;+,,+$+$!
K?c>_+$2!@0(!DJ`H!59!R+$)&+567!"#!-dV/!DJZF!+$"5+&2!4+$*+(E!a7(&567+!K?c>_+$2+!4#$>
*+(! 8Q$! *5+! O0824'$+! -dV/! ;+$+52,! M#;&5"5+$2! xc'9;$5*)+! c$N,2'&&0)$'M756! /'2'! c+(2$+n!
722Mzoo444E66*6E6'9E'6E#1oyE!
!PDPDP! Q#'*2.99.+!Screeening!%),-!Y$*.%*#.99.%!/%-#;#*$'.%!0.'!B*IF=!2%0!0.'!>&IF=!
:Q$!*',!U067*#$67,'2">Docking!952!*+$!O0824'$+!LVK3[O!4#$*+(!"#@0$!W5(',+>X(75;520$+(!
'#,!*+$!<5(*5()/<!,+&+125+$2J!*5+!+5(+(!XcZD>_+$2!@0(!#(2+$!GD!ÜB!"+5)2+(E!/5+,+$!O67$522!
4#$*+!*#$67)+8Q7$2J!#9!+5(+(![+,2*'2+(,'2"!)++5)(+2+$! X(75;520$+(!7#9'(+$!W5(',+(!"#!+$>
7'&2+(E!/5+,+!X(75;520$+(!,0&&2+(!8Q$!M'$',52A$+!^5+&M$02+5(+!0M2595+$2!4+$*+(!1g((+(J!*'!*5+!
K[L><5(*+2',67+(! 7#9'(+$! #(*! M'$',52A$+$! W5(',+(! ,+7$! A7(&567! ,5(*E! X9!U067*$#67,'2">
Docking! 4#$*+(! 5(,)+,'92! CEGYH!W5(',+>X(75;520$+(! *+$! <5(*5()/<! O#;,2'("*'2+(;'(1! 5(!
*5+!'125@+(!^+(2$+(!*+$!U090&0)5+90*+&&+!%2BPW!#(*!L8BPW!+5()+M',,2!#(*!9522+&,!*+$!
LVK3[O>Scoringfunktion!;+4+$2+2E!!!
B52!*+9!U090&0)5+90*+&&!*+$!%2BPW!10((2+(!59!@5$2#+&&+(!Screening!7'#M2,A67&567!X(75;5>
20$+(! *+$! 7#9'(+(!c/WI! #(*! *+$! 7#9'(+(! L$02+5(!W5(',+!<! eLW<f! '()+$+567+$2!4+$*+(E!
^#*+9!8'(*!+5(+!K($+567+$#()!@0(!X(75;520$+(!*+$!7#9'(+(!c/WGJ!c/WC!#(*!c/W]!,2'22E!
X(!K;;5&*#()!YEGF!,5(*!*5+!K($+567+$#(),8'120$+(!e%:!{!Enrichment Factorf!*+$!;+1'((2+(!
W5(',+>X(75;520$+(! 5(! *+(! 0;+$+(! Gq! e%:GfJ! Cq! e%:Cf! ;"4E! Zq! e%:Zf! *+$! 85('&+(! 952!
LVK3[O>Score!;+4+$2+2+(!/'2+(;'(1!*'$)+,2+&&2E!X9!:'&&!*+$!L8BPW!10((2+(!7'#M2,A67&567!
X(75;520$+(!*+$!Epidermal-Growth-Factor-Receptor!W5(',+!e%-:>PfJ!*+$!Y>L70,M705(0,525*+>
';7A()5)+(!L$02+5(!W5(',+!G! eL/WGf!#(*!*+$!LW<! 5(!*+(!0;+$+(! "4+5!L$0"+(2!*+$! 85('&+(!
/'2+(;'(1!'()+$+567+$2!4+$*+(E! X(!*+(!0;+$+(!Z!L$0"+(2!*+$!/'2+(;'(1!8'(*+(!,567!"#*+9!
'#67! X(75;520$+(! *+$! [7N95*5(>W5(',+! #(*! c'Ctoc'&90*#&5(>*+M+(*+(2! L$02+5(! W5(',+!
ecKBWfE!!
! ! ERGEBNISSE
FI!
W[WW
E[WW
2[WW
C[WW
Y[WW
)W[WW
)E[WW
)2[WW
)C[WW
)Y[WW
0>7A75(
!'()
!'(E
!'(2
!'(U
!;6)
1I#<T
b(0
b(/
b(!
I1.)6VW).*12(U$,#/.
1#)
1#E
1#P
!
Abbildung 3.18: Anreicherungen der jeweiligen Kinase-Inhibitoren für die EtMRK.
/5+!<+4+$2#(),8#(1250(!LVK3[O>Score!4#$*+!8Q$!*',!85('&+!Ranking!*+$!/'2+(;'(1!@+$4+(*+2E!K#8!*+$!d$*5>
('2+!,5(*!*5+!p+4+5&5)+(!W5(',+>X(75;520$+(!'#8)+2$')+(E!/5+!K;,"5,,+!;+"+567(+2!*5+!p+4+5&5)+(!K($+567+$#(),>
8'120$+(!e%:fJ!5(!*+(!0;+$+(!Gq!e%:GfJ!Cq!e%:Cf!#(*!Zq!e%:Zf!*+$!85('&+(!/'2+(;'(1E!!!
W[WW
E[WW
2[WW
C[WW
Y[WW
)W[WW
)E[WW
)2[WW
)C[WW
)Y[WW
0>7A75(
!0f(
!'()
!'(E
!'(2
IQ(S
1I#<T
b'()
b(0
b(/
b(!
H;+F-?-.8(
I1.)6VW).*12(U$,#/.
1#)
1#E
1#P
!!!
Abbildung 3.19: Anreicherungen für die PfMRK der jeweiligen Kinase-Inhibitoren.
/5+!<+4+$2#(),8#(1250(!LVK3[O>Score!4#$*+!8Q$!*',!85('&+!Ranking!*+$!/'2+(;'(1!@+$4+(*+2E!/5+!K;,"5,,+!
;+"+567(+2!*5+!p+4+5&5)+!K($+567+$#()J!5(!*+(!0;+$+(!Gq!e%:GfJ!Cq!e%:Cf!#(*!Zq!e%:Zf!*+$!85('&+(!/'2+(;'(1E!
ERGEBNISSE
FF!
!
?9!)++5)(+2+!O2$#12#$1&',,+(!'&,!@5$2#+&&+!Hits!'#,"#4A7&+(J!4#$*+!8Q$!*5+!"4+5!L$0"+(2!*+$!
O2$#12#$+(!952!*+9!7g67,2+(!LVK3[O>Score!*5@+$,+!Murcko Assemblies!x<%BXO!k!B?P>
cWdJ!GHH`y!)+(+$5+$2E!/5+!,0!,+&+125+$2+(!O#;,2'("+(!*5+(2+(!'&,!Hit-Cluster!*+$!M'$',52A$+(!
^5+&M$02+5(+J!*5+!)+)+;+(+(8'&&,! 59!K(,67&#,,!(067!4+52+$!0M2595+$2!4+$*+(!1g((+(J! ,08+$(!
,567!57$+!K125@52A2!in vitro!;+,2A25)+(!&A,,2E!:Q$!p+*+,!^5+&M$02+5(!4#$*+(!'672!O2$#12#$+(!'#,>
)+4A7&2E!/$+5!*5+,+$!'672!O2$#12#$+(!4#$*+(!,0407&!8Q$!*5+!%2cPWC!'&,!'#67!8Q$!*5+!L8BPW!
5*+(2585"5+$2!e,5+7+
! ! DISKUSSION
FH!
S!A#+52++#$%!!
SDH! 7859#%:);-<%3#3.!=#%)+.%!)9+!7#.9Y'$*.#%.!0.'!M-.')Y#.!6$%!>'$*$:1$.%:/%&.5*#$%.%!
^N1&5(>';7A()5)+!W5(',+(!,5(*!^5+&M$02+5(+!5(!*+$!<+1A9M8#()!@0(!W$+;,+$1$'(1#()+(J!*'!
,5+! +5(+! O67&Q,,+&8#(1250(! 5(! *+$! P+)#&'250(! *+,! ^+&&"N1&#,! ;+,52"+(! xBKV?B<P%O! k!
<KP<KcX/J!CDDHyE!/+$!$+5;#(),&0,+!K;&'#8!*+,!^+&&"N1&#,!5,2!8Q$!p+*+!,567!2+5&+(*+!^+&&+!l!
'#67!8Q$![#90$"+&&+(!l!+,,+(25+&&!x=dUK3OOd3!k!L%POOd3J!CDDFyE!X(![#90$"+&&+(!5,2!*+$!
^+&&"N1&#,! *+$+)#&5+$2! x<%P[U%[! k! WKV/XOJ! CDDIyE! /5+! :0&)+! 5,2! +5(! #(10(2$0&&5+$2+,!
^+&&4'67,2#9!x/%V!OKVJ!GHHIyE!%5(+!)+"5+&2+!X(75;5+$#()!@0(!^+&&"N1&#,>$+&+@'(2+(!c/W,!
#(2+$;5(*+2!+5(!:0$2,67$+52+(!*+$![#90$;5&*#()!#(*!7'2!(#$!)+$5()+(!%5(8&#,,!'#8!$#7+(*+,!l!
(5672! M$0&58+$5+$+(*+,! -+4+;+! x<%P[U%[! k! WKV/XOJ! CDDIn! Bd3Kcd! k! RKVVK3dJ!
CDDYyE! W$+;,"+&&+(! #(*! M'$',525,67! &+;+(*+! L$020"0+(! A7(+&(! +5('(*+$! xWVX3W%P[! k!
U%?OOV%PJ!CDD`yE!<+5*+!;+,52"+(!+5(+!@+$)&+567;'$!707+!^+&&2+5&#(),$'2+!x/d%PX-J!CDD]yE!
/5+,+!-+9+5(,'91+52!5(,M5$5+$2+!*5+!K#,4'7&!@0(!^N1&5(>';7A()5)+(!W5(',+(!'&,!^5+&M$02+5>
(+! 5(! M'$',525,67! &+;+(*+(!L$020"0+(E!/5+!%5)(#()! *5+,+$!^5+&M$02+5(+! 10((2+! 5(!L$020"0+(!
;+$+52,!9+7$8'67!)+"+5)2!4+$*+(!x/d%PX-J!CDD]n!UKBBKP[d3 et al.J!CDDYn!3K?VK et
al.J!CDDZyE!K('&0)!,0&&2+!*5+!X(75;5+$#()!@0(!c/W,!'#,!E. tenella!*+(!^+&&"N1&#,!#(2+$;5(*+(!
#(*!*'952!*+(!V+;+(,"N1&#,!*+,!L'$',52+(!#(2+$;$+67+(E!X(!in vitro!%\M+$59+(2+(!10((2+!*5+,!
('67)+45+,+(!4+$*+(E!U5+$8Q$!4#$*+!*+$!%88+12!*+,!;+1'((2+(!c/W>X(75;520$,!:&'@0M5$5*0&!
'#8! *5+! X(75;5+$#()! *+$!O675"0(2+(+(24561&#()!)+2+,2+2! e,5+7+!K;,67(522! GEZEZJ
DISKUSSION
HD!
%2cPWC!*$+5!4+52+$+!c/W>A7(&567+!O+b#+("+(!5*+(2585"5+$2E!X9!B+(,67+(!,5(*!+&8!c/W!L$0>
2+5(+!;+1'((2E!U5+$@0(!;+,52"+(!@5+$!c/W!L$02+5(+!+5(+!456725)+!:#(1250(!5(!*+$!P+)#&'250(!
*+,!^+&&"N1&#,!eU,c/WGJ!U,c/WCJ!U,c/W]!#(*!U,c/W`fE!:Q$!U+8+(!5,2!;+1'((2J!*',,!*5+!
^+&&"N1&#,$+)#&'250(! (#$! *#$67! +5(+! +5("5)+! c/W! e6*6CMoc/cCF! +b#5@'&+(2! "#$! 7#9'(+(!
c/WGf!952!@+$,675+*+(+(!^N1&5(M'$2(+$(!)+,2+#+$2!45$*!xBdP-K3J!GHHIyE!%,!5,2!,0952!'(>
"#(+79+(J!*',,!*5+!c/W!L$02+5(+!5(!X(@+$2+;$'2+(!9+7$+$+!K#8)';+(!5(!*+$!^+&&"N1&#,$+)#>
&'250(!Q;+$(+79+(J!#(*!*',,!;+5!+5(+9!:#(1250(,@+$&#,2!)$'@5+$+(*+!:0&)+(!8Q$!*+(!d$)'(5,>
9#,!+(2,2+7+(E!/#$67!O+b#+("@+$)&+567+!10((2+(!5(!E. tenella!(#$!"4+5!c/W!L$02+5(+!5*+(25>
85"5+$2!4+$*+(!e%2cPWC!#(*!%2cPWYfJ!*5+!@+$9#2&567!+5(+!P0&&+!59!^+&&"N1&#,!,M5+&+(E!U5+$>
;+5!5,2!%2cPWC!7090&0)!"#$!U,c/WG!,045+!U,c/WCE!/')+)+(!;+,52"2!%2cPWY!*5+!7g67,2+!
X*+(252A2!"#$!U,c/WGE!/5+!R+$9#2#()J!*',,!*5+!%2cPWC!+5(+!"+(2$'&+!:#(1250(!59!^+&&"N1>
&#,!@0(!E. tenella!;+,52"2J!45$*!*#$67!R+$,#67+!952!'(*+$+(!c/W!7090&0)+(!L$02+5(+(!M'$'>
,52A$+$!L$020"0+(!)+,2Q2"2! x/d%PX- et al.J! CDDDyE!%,!10((2+!)+"+5)2!4+$*+(J! *',,! *5+!R+$>
9+7$#()!*+$!L'$',52+(!Plasmodium falciparum,!Leishmania mexicana!#(*!Trypanosoma bru-
cei! *#$67!c/W>X(75;520$+(J! *5+! 8Q$! 7#9'(+!c/W,! +(24561+&2!4#$*+(J! #(2+$;#(*+(!4+$*+(!
10((2+!x-PK3[ et al.J!CDD]n!UKPBO% et al.J!CDDGn!_dd/KP/ et al.J!CDDYyE!!!
/5+!<VKO[>O#67+!5(!/'2+(;'(1+(!*+,!O'()+$!X(,252#2+,J!+$)';!*5+!;5,&'()!(5672!;+,67$5+;+>
(+(!O+b#+("+(!%2BPWJ!%2cPWG!#(*!%2cPWYE!/',,!+,!,567!;+5!*+(!)+8#(*+(+(!O+b#+("+(!
2'2,A67&567!#9!^N1&5(>';7A()5)+!W5(',+(!7'(*+&2J!;+,2A25)2!*5+!707+!O+b#+("5*+(252A2!,0407&!
"#!%2cPWC!'&,!'#67!"#!L8LWZ!,045+!"#!'(*+$+(!7090&0)+(!c/W>O+b#+("+(!@0(!P. falcipa-
rumE! %5(!9#&25M&+,! O+b#+(">Alignment! 952! *+$! 7#9'(+(! c/WC! O+b#+("! "+5)2+! '#r+$*+9J!
*',,!'&&+!*$+5!O+b#+("+(!*5+!10(,+$@5+$2+(!/09A(+(!->LoopJ!LO[KXP%>U+&5\J!Hinge>P+)50(!
#(*![>Loop!;+,52"+(E!/5+!O+b#+("+(!4'$+(!8Q$!%2cPWY!,045+!8Q$!%2cPWG!'(!;+5*+(![+$95>
(5!#(@0&&,2A(*5)J!,0!*',,!8Q$!;+5*+!O+b#+("+(!1+5(+!@0&&,2A(*5)+(!L$02+5(,+b#+("+(!';)+&+52+2!
4+$*+(!10((2+(E!:Q$!*5+!%2BPW!10((2+! p+*067!+5(+!@0&&,2A(*5)+!L$02+5(,+b#+("!@0$7+$)+>
,')2!4+$*+(E!O5+!45+,!+5(+!)$0r+!3>2+$95('&+!R+$&A()+$#()!@0(!]Y!K95(0,A#$+(!'#8E!K#8>
)$#(*!*+$
! ! DISKUSSION
HG!
7'&;!*+$!ApicomplexaE!%5(+!+\M+$59+(2+&&+!L$Q8#()!*5+,+$!UNM027+,+!4#$*+!(5672!*#$67)+>
8Q7$2E!!!
%5(+!R0$7+$,')+!*+$!:#(1250(J!*+$! 59!P'79+(!*5+,+$!K$;+52! 5*+(2585"5+$2+(!c/W>A7(&567+(!
L$02+5(+!@0(!E. tenellaJ! 5,2!9522+&,!O+b#+("@+$)&+567!"#!O+b#+("+(!)#2!67'$'12+$5,5+$2+$!7#>
9'(+$!c/W,!(5672!@0&&,2A(*5)!9g)&567E!%2BPW!;+,52"2!+5(+!707+!O+b#+("5*+(252A2!"#$!7#9'>
(+(!c/WI!eU,c/WIfE!/'7+$!4A$+!+5(+!0$270&0)+!:#(1250(!"#$!U,c/WI!*+(1;'$E!:Q$!*5+,+!
5,2! +5(+! *#'&+! :#(1250(! ;+,2A25)2! 40$*+(! x:XOU%PJ! CDDZyE! U,c/WI! 5,2! "#9! +5(+(! '(! *+$!
[$'(,1$5M250(,$+)#&'250(! *+$!P3K>L0&N9+$',+! XX! ;+2+5&5)2! #(*! "#9!'(*+$+(! ;+,52"2! ,5+! +5(+!
:#(1250(!'&,!c/W>K125@5+$#(),15(',+!ecKWfE!/5+!O+b#+("'('&N,+!*+$!%2BPW!&5+8+$2+!U5(>
4+5,+J! *5+! )+)+(! +5(+! "#$! U,c/WI! 0$270&0)+! :#(1250(! ,M$+67+(E! %,! 10((2+! 1+5(! W+$(>
V01'&5,5+$#(),,5)('&! e3VOf! xB%X%P!k!<Vd<%VJ!GHHDy! 5(!*+$!%2BPW!O+b#+("!@0$7+$)+>
,')2!4+$*+(E!L$02+5(+!*5+!'(!*+$![$'(,1$5M250(,$+)#&'250(!;+2+5&5)2!,5(*J!9Q,,+(!5(!*+(!W+$(!
2$'(,M0$25+$2!4+$*+(!#(*!;+(g25)+(!75+$8Q$! +5(!3VOJ! *',! *+(![$'(,M0$2! 5(! *+(!^+&&1+$(!)+>
4A7$&+5,2+2!xVK3-% et al.J!CDDHyE!%5(+!cKW>:#(1250(!*+$!%2BPW!5,2!'#8)$#(*!*+,!:+7&+(,!
@0(!cKW><5(*+9025@+(!5(!*+(!M02+(25+&&+(!O#;,2$'2+(!%2cPWC!#(*!%2cPWY!+;+(8'&&,!8$')>
&567E! ^#$! @0&&,2A(*5)+(!K125@5+$#()! *+$! %2cPWC! ,045+! *+$! %2cPWY!9#,,! *5+!K95(0,A#$+!
[7$+0(5(!59!K125@5+$#(),>Loop!*5+,+$!W5(',+(!M70,M70$N&5+$2!4+$*+(E!/5+,!1'((!'#67!*#$67!
+5(+! K#20M70,M70$N&5+$#()! +$80&)+(! #(*! 4#$*+! 8Q$! *5+! %2cPWC! ;+$+52,! ('67)+45+,+(!
xWX33KXP/ et al.J!CDD]yE!/'7+$!4A$+!+5(!cKW>%("N9!"#$!@0&&,2A(*5)+(!K125@5+$#()!(5672!
"45()+(*! (024+(*5)E! /5+,+! R+$9#2#()! 45$*! *'*#$67! #(2+$,2Q2"2J! *',,! 4+*+$! *5+! %2cPWC!
(067!*5+!%2cPWY!+5(!cKW><5(*+9025@!;+,52"+(E!!!
/5+!%2cPWY!L$02+5(,+b#+("! 5,2!*+$!L$02+5(,+b#+("!*+$!U,c/WG!A7(&567E!/5+!X('125@5+$#()!
*+$!U,c/WG!45$*!*#$67!L70,M70$N&5+$#()!'(!*+(!K95(0,A#$+(![7$G]!,045+![N$GZ!$+)#&5+$2!
#(*! 5,2! 8Q$! *+(! c/WGo^N1&5(>W09M&+\! 67'$'12+$5,25,67! xBKV?B<P%O! k! <KP<KcX/J!
CDDZyE!_A7$+(*![N$0,5(! 5(! '&&+(!cPWY>A7(&567+(! L$02+5(,+b#+("+(! *+$!Apicomplexa 10(>
,+$@5+$2!@0$&5+)2J!&5+)2!'(!*+$!7090&0)+(!L0,5250(!*+,![7$G]!+5(!K&'(5(!@0$E!/5+,!&A,,2!@+$9#>
2+(J!*',,!*5+!9g)&567+!X('125@5+$#()!*#$67!+5(+(!;5,7+$!(067!#(;+1'((2+(!B+67'(5,9#,!$+>
)#&5+$2!45$*E!
DISKUSSION
HC!
SDP! "#$#%&$'()*#+,-.!/0.%*##.'2%3!2%0!4-)')5*.'#+#.'2%3!6$%!7859#%.%!#%!E. tenella!
%5(!4+52+$+,!^5+&!*5+,+$!K$;+52!4'$!*5+!X*+(2585"5+$#()!^N1&5(>A7(&567+$!O+b#+("+(!5(!E. te-
nellaE!/5+!K#,4'7&!*+$!O#67,+b#+("+(!8Q$!*5+!<VKO[>O#67+!('67!(5672!5*+(2585"5+$2+(!^N1&5>
(+(J!*5+!9g)&567+$4+5,+!5(!*+$!V')+!,5(*J!*5+!^N1&5(>';7A()5)+!W5(',+!%2cPWC!'#,!E. tenel-
la!"#!'125@5+$+(J!+$80&)2+!'#8!<',5,!;+1'((2+$!^N1&5(+!'#,!P. falciparum!eL8cjcGJ!L8cjcCJ!
L8cjcY!#(*!L8cjc]f!xB%PcW} et al.J!CDDYyE!_5+!B%PcW}!et alE!"+5)+(!10((2+(J!,5(*!
*5+!L$02+5(+!L8cjcG!#(*!L8cjcY!5(!*+$!V')+J!*',,!L8LWZ!L$02+5(!"#!'125@5+$+(!xB%PcW}
et al.J! CDDYyE!K('&0)! "#$!O52#'250(! 5(!P. falciparum! 5,2!%2cjcG! ,045+!%2cjcY!@+$9#2&567!
+;+(8'&&,! )++5)(+2! %2cPWC! "#! '125@5+$+(E!/'$Q;+$! 75('#,! 5,2! '#67!PX3-d! x:%P<j et al.J!
GHHHyJ!+5(!L$02+5(!'#,!Xenopus laevisJ!*',!1+5(+!O+b#+("A7(&5671+52!952!*+(!^N1&5(+(!;+,52"2J!
p+*067! ,2$#12#$+&&+!a7(&5671+52+(!'#84+5,2J! 5(!*+$!V')+J!L8LWZ!"#!'125@5+$+(! xB%PcW} et
al.J! CDDYyE!B'$�'!VE! O#Ä$+"! :+$(Ä(*+"! eX(2+$@+2! X((0@'250(!-9;UJ! O674';+(7+59f! 10((2+!
"+5)+(J!*',,!PX3-d!)++5)(+2! 5,2J!*5+!%2cPWC!"#!'125@5+$+(E!/5+!952!*+$!<VKO[>O#67+!"#!
L8cjcG!#(*!L8cjcY!)+8#(*+(+(!^N1&5(+!e%2cjcGJ!%2cjcY'!,045+!%2cjcY;fJ!,0&&2+(!'#,!
*5+,+9!-$#(*!+;+(8'&&,!5(!*+$!V')+!,+5(J!*5+!'(*+$+(!c/W>7090&0)+(!L$02+5(+!"#!'125@5+$+(E!
/5+,! 10((2+! @0(!B'$�'!VE! O#Ä$+"!:+$(Ä(*+"! in vitro! 8Q$! *5+!%2cjcY'!)+"+5)2!4+$*+(! #(*!
;+,2A25)2! ,0! *5+! '()+(099+(+
! ! DISKUSSION
HY!
/5+!952! *+$! O#67,+b#+("! L8cjcY! *#$67)+8Q7$2+! <VKO[>O#67+! +$)';! "4+5! 7090&0)+! O+>
b#+("+(!5(!E. tenella e%2cjcY'!#(*!%2cjcY;fE!%2cjcY'J!%2cjcY;!,045+!L8cjcY!;+,52>
"+(!*5+!7g67,2+!O+b#+("A7(&5671+52!"#![)cjcC!'#,!T. gondii!xUKBBKP[d3 et al.J!CDDYyE!
[)cjcC!"+5)2!1+5(+!O+b#+("A7(&5671+52!"#!*+(!7#9'(+(!-G>L7',+!$+)#&5+$+(*+(!^N1&5(+(!
/!#(*!%J!5,2!';+$!5(!*+$!V')+J!*5+!59!^+&&"N1&#,!$+&+@'(2+![)cPWY!"#!'125@5+$+(E!['cPWY!5,2!
7090&0)! "#! %2cPWCE! /'7+$! &5+)2! *5+! R+$9#2#()! ('7+J! *',,! '#67! %2cjcY'! '&,! '#67! %2>
cjcY;!)++5)(+2!,5(*J!%2cPWC!"#!'125@5+$+(E!_5+!;+$+52,!;+,67$5+;+(J!10((2+!*5+,+![7+,+!8Q$!
*5+!%2cjcY;!in vitro!;+,2A25)2!4+$*+(E!!
/5+!K;8$')+,+b#+("+(!L8cjcC!#(*!L8cjc]! ,5(*!;5,&'()!(5672!*+2'5&&5+$2! ;+,67$5+;+(E!:Q$!
L8cjcC!5,2!*5+!)+('#+!:#(1250(!;5,&'()!(5672!;+1'((2!#(*!+,!10((2+!'#67!1+5(+!7090&0)+!
O+b#+("!59!-+(09!@0(!E. tenella!5*+(2585"5+$2!4+$*+(E!:Q$!L8cjc]!@+$9#2+2!9'(!+5(+!:#(1>
250(! '&,! K125@5+$#(),M'$2(+$! *+$! L8cPWG! 5(! P. falciparumE! X(! E. tenella! 10((2+! +5(! "#!
L8cPWG!7090&0)+,!L$02+5(!('67)+45+,+(!4+$*+(!e%2cPWGfE!K('&0)!"#!P. falciparum!1'((!
*'7+$!@+$9#2&567!*5+!%2cPWG!+;+(8'&&,!*#$67!*',!^N1&5(!%2cjc]!'125@5+$2!4+$*+(E!!!
X(! +#1'$N025,67+(!ON,2+9+(! 5,2! *5+!W0(2$0&&+!*+,!^+&&"N1&#,! ,+7$! )#2! #(2+$,#672E!%,! 5,2! ;+>
1'((2J!*',,!*5+!K("'7&!*+$!^N1&5(+!4+,+(2&567!7g7+$!5,2J!'&,!*5+!K("'7&!*+$!1'2'&N25,67+(!?(>
2+$+5(7+52+(! xBKV?B<P%O! k! <KP<KcX/J! CDDZn! OK[jK3KPKjK3K! k! WKV/XOJ!
CDDHyE!%,!4#$*+!'#67!('67)+45+,+(J!*',,!*5+,+!@0(!@+$,675+*+(+(!^N1&5(+(!'125@5+$2!4+$*+(!
xUdcU%--%P et al.J! CDDFn!BKV?B<P%OJ! CDDZyE! X(!P. falciparum! 75()+)+(!4#$*+! (#$!
+5(+!*+#2&567!)+$5()+$+!K("'7&!@0(!^N1&5(+(!,045+!c/W,!)+8#(*+(!x/d%PX- et al.J!CDDCn!
B%PcW} et al.J!CDDYyE!X(!*5+,+$!K$;+52! 5,2!+,!*#$67! in silico!B+270*+(!)+&#()+(J!*$+5!;5,>
&'()!(5672!'((025+$2+!c/W>A7(&567+!L$02+5(+!#(*!@5+$!^N1&5(+!5(!E. tenella!"#!5*+(2585"5+$+(E!
%,!4#$*+!,0952!+5(+!@+$)&+567;'$+!K("'7&!@0(!c/W,!,045+!K125@'20$+(!45+!5(!'(*+$+(!d$>
)'(5,9+(!*+$!Apicomplexa!5*+(2585"5+$2E!%,!5,2!*+(1;'$J!*',,!(067!4+52+$+!7090&0)+!L$02+5(+!
5(!E. tenella!+\5,25+$+(J!*'!9522+&,!Best-Best-Hit!;+5!+5(+$!<VKO[>O#67+!(5672!"4'(),&A#85)!
'&&+!U090&0)+(!5*+(2585"5+$2!4+$*+(E!/5+,!&5+)2!*'$5(!;+)$Q(*+2J!*',,!*+$!;+,2+
DISKUSSION
H]!
falciparum!;+$+52,!*#$67)+8Q7$2!x/d%PX- et al.J!CDDCn!B%PcW} et al.J!CDDYy!l!*#$67!;50>
67+95,67+!%\M+$59+(2+!;+&+)2!4+$*+(E!!
SDS! G*')*.3#.%!12'!X.((2%3!6$%!=#%)+.%!
:Q$! +5(+!U+99#()! @0(!W5(',+(! ;5+2+(! ,567! )$#(*,A2"&567! @5+$! @+$,675+*+(+!O2$'2+)5+(! '(z!
+5(+!<&0615+$#()!*+$!O#;,2$'2;5(*#()J!*5+!R+$*$A()#()!@0(!K[LJ!+5(+!'&&0,2+$5,67+!P+)#&'>
250(!,045+!+5(+!X$$+@+$,5;&+!X(75;5+$#()!xW?<X3jX!k!BsVV%PJ!CDD]yE!X9!+$,2+(!:'&&!9#,,!
*5+!K#,;5&*#()!+5(+,!L$02+5(>L$02+5(>W0(2'12+,!@+$75(*+$2!4+$*+(J!*'!W5(',+(!'(*+$+!L$02+5>
(+!'&,!O#;,2$'2+!+$1+((+(!#(*!;5(*+(!xWV%<%J!CDDHyE!K#8)$#(*!*+$!-$gr+!*+$!,567!'#,;5&>
*+(*+(!_+67,+&45$1#(),8&A67+(!+$,67+5(2!*5+,+!O2$'2+)5+!A#r+$,2!,6745+$5)J! p+*067!,5(*!+5>
(5)+!<+5,M5+&+!'#,!*+$!V52+$'2#$!;+1'((2J!5(!*+(+(!X(75;520$+(!*5+!O#;,2$'2;5(*+2',67+!'*$+,>
,5+$+(!xdP^K%^ et al.J!CDDHyE!/+$!R0$2+5&!+5(+$!<&0615+$#()!*+$!O#;,2$'2;5(*#()!&5+)2!5(!*+$!
)+$5()+(!W0(,+$@5+$#()!59!<+$+567!*+$!O#;,2$'2><5(*+2',67+J!4',!+,!+$9g)&5672J!O+&+125@52A2!
"#!+$$+567+(!xW?<X3jX!k!BsVV%PJ!CDD]yE!%5(+!R+$*$A()#()!*+,!K[L!'#,!,+5(+$!<5(*+2'>
,67+! ;+5,M5+&,4+5,+! 8Q7$2! '#8)$#(*! *+$! 707+(!W0(,+$@5+$#()! 7A#85)! "#! O+&+125@52A2,M$0;&+>
9+(J!#(*!1g((2+!,0952!#9)'()+(!4+$*+(!xdP^K%^ et al.J!CDDHyE!-+$5()8Q)5)+!?(2+$,675+*+!
*+$!K[L><5(*+2',67+(!@0(!c/W,!*+$!R+$2+;$'2+(!#(*!*+$!L$020"0+(!e'#,8Q7$&567+$!*5,1#25+$2!
5(!K;,67(522!]EZfJ!+$&'#;+(!p+*067!*5+!%(24561&#()!@0(!,+&+125@+(!K[L>109M+2525@+(!X(75;5>
20$+(E!!
SDT! V.#*+*2'5*2'#0.%*##.'2%3!6$%!/%-#;#*$'.%!0.'!B*4F=J!
:Q$!*5+!O#67+!('67!(+#+(!X(75;520$+(!*5+!*',!%2cPWC!L$02+5(!'*$+,,5+$+(!4#$*+!;+$+52,!+5(!
virtuelles Screening! *#$67)+8Q7$2! x<%j%PJ! CDDIyE! /',! @0(! <+N+$! @+$4+(*+2+2! Screening>
L$02010&&! 5,2! A7(&567! "#! @5+&+(! '(*+$+(! O2#*5+(! xB?[UKO et al.J! CDDFn! d%VVX%3 et al.J!
CDDHn! L%PdVKJ! CDD`yE! %,! #98',,2+! +5(+! Y/>L7'$9'10M70$85&2+$#()J!90&+1#&'$+,!Docking!
#(*! Scoring! ,045+! @+$,675+*+(+! 3'67M$0"+,,5+$#(),,67$522+E! U5+$;+5! 10((2+(! +&8! %2cPWC>
X(75;520$+(!5*+(2585"5+$2!4+$*+(!x<%j%PJ!CDDIyE!!!
%5(! 2NM5,67+$!O67$522! 8Q$! *',! ,567! '(,67&5+r+(*+! $'250('&+!_5$1,2088*+,5)(! 5,2! *5+!K($+567+>
$#()!@0(!Hit-Clustern!xOcU3%cW%!k!<dO[PdBJ!CDD`yE!U5+$"#!4#$*+(!O#;,2$#12#$>!#(*!
a7(&5671+52,,#67+(! '#8!<',5,! *5+,+$! X(75;520$+(! *#$67)+8Q7$2E!/5+! +5()+,+2"2+!B+270*+! ;'>
,5+$2!'#8!*+$!K(('79+J!*',,!,2$#12#$+&&!A7(&567+!R+$;5(*#()+(!'#67!+5(+!A7(&567+!;50&0)5,67+!
K125@52A2!"+5)+(!xi?X3[?O et al.J!CDDHyE!K&,!O67&Q,,+&+5)+(,67'82!8Q$!*5+!O#;,2$#12#$,#67+(!
! ! DISKUSSION
HZ!
4#$*+!*+$ Scaffold!5(!*+(!;+$+52,!'125@+(!O#;,2'("+(!;+,25992J!*+$!'(!*5+!Hinge>P+)50(!;5(>
*+2!#(*!,0952!+,,+(25+&&!8Q$!*5+!X(75;520$;5(*#()!5,2E!/#$67!Dockin)>%\M+$59+(2+!4#$*+(!*5+!
,0! 5*+(2585"5+$2+(! O#;,2'("+(! ;+4+$2+2! #(*! '#,)+4A7&2E! :Q$! ,+67,! @0(! GYH! @5$2#+&&+(!Hits!
10((2+! in vitro!K125@52A2! ;+,2A25)2!4+$*+(E! X9!109;5(5+$2+(! in silico! #(*! in vitro!Workflow!
10((2+(!5(,)+,'92!@5+$!M02+(25+&&+!V+52,2$#12#$+(!*+$!%2cPWC!5*+(2585"5+$2!4+$*+(E!/5+!5(2+>
$+,,'(2+,2+!W&',,+!*+$!%2cPWC>X(75;520$+(!,5(*!*5+!<+("595*'"0&>c'$;0(52$5&+J!*'!,5+!;5,&'()!
5(!*+$!V52+$'2#$!(#$!@0(!<'*'4+NJ!W'MM+!#(*!%&!U'4',7!x<K/K_%j!k!WKLL%J!GHHHn!%V!
UK_KOU et al.J!GHHHy!,045+!5(!+5(+9!L'2+(2!xKP<XO%P!k!KB<VKP/J!CDD`y!5(!^#,'9>
9+(7'()!952!W5(',+(!;+,67$5+;+(!4#$*+(E!K#,!*5+,+9!-$#(*!4#$*+!*5+,+! in silico!4+52+$!
0M2595+$2!#(*!O2$#12#$+(!8Q$!+5(+!67+95,67+!ON(27+,+!@0$)+,67&')+(E!K&&+!'(*+$+(!O2$#12#$>
1&',,+(!e3'M720&'62'9+J!P70*'(5(!#(*!d\5(*0&+f!,5(*!,670(!&A()+$+!^+52!'&,!X(75;520$+(!*+$!
c/W,!0*+$!-&N60)+(,N(27',+(!e-OWf!;+1'((2!x<PKBOd3 et al.J!CDDGn!-PKj et al.J!GHHHn!
VX? et al.J! CDDYyE! /5+! :'95&5+(! *+$! c/W! #(*! *+$! -OW! ,5(*! +()! 952+5('(*+$! @+$4'(*2!
xBK33X3- et al.J! CDDCyJ! *'7+$! 5,2! +,! ('7+&5+)+(*J! *',,! A7(&567+! O2$#12#$2NM+(! 5(! ;+5*+(!
L$02+5(8'95&5+(!+5(+!K125@52A2!"+5)+(E!/5+!in silico!B+270*+(!4'$+(!,0952!5(!*+$!V')+J!5(2+>
$+,,'(2+!R+$;5(*#()+(!"#!5*+(2585"5+$+(E!!
W+5(+!*+$!@5+$!5*+(2585"5+$2+(!V+52,2$#12#$+(!*+$!%2cPWC!"+5)2+!O+&+125@52A2!8Q$!*5+!M'$',52A$+!
W5(',+E!/5+!%$)+;(5,,+!*+$!;5067+95,67+(!%\M+$59+(2+!"+5)+(!p+*067!*+#2&567+!?(2+$,675+*+!
5(!*+$!K125@52A2!)+)+(Q;+$!U,c/WC!#(*!%2cPWCE!/5+!V+52,2$#12#$+(!45+,+(!(5+*$5)+$+!XcZD>
_+$2+! #(*! W5>_+$2+! '#8! *5+! U,c/WC! '&,! '#8! *5+! %2cPWC! '#8! e,5+7+! "E<E! 3'M720&'62'9J!
[';+&&+!YEIJ![';+&&+!YEF! 59!K;,67(522!YECEYfE!/5+,+!%$)+;(5,,+!)+;+(!+5(+(!U5(4+5,!*'$'#8J!
*',,! +,! ,5)(5851'(2+!?(2+$,675+*+! "45,67+(! *+$!U,c/WC! ,045+! *+$!-)c/WY! #(*! M'$',525>
,67+(!c/WC!L$02+5(+(!)+;+(!9#,,J!#(*!*5+,+!,0952!'#67!@0(!X(75;520$+(!'*$+,,5+$2!4+$*+(!
1g((+(E!!!
d;407&! c/W! L$02+5(+! @0(! OA#)+25+$+(! +5(+! 707+! O+b#+("5*+(252A2! "#+5('(*+$! '#84+5,+(J!
10((2+(! ,+&+125@+! X(75;520$+(! 8Q$! +5("+&(+! c/W>O#;1&',,+(! 5*+(2585"5+$2! 4+$*+(!
xWX[K-K_K et al.J!GHHYn!B%X=%P et al.J!GHHIyE!U5+$;+5!1'((!+5(!+5("+&(+$!K95(0,A#$+'#,>
2'#,67!'#,$+567+(J!*'952!+5(!X(75;520$!,+&+125@!+5(+!c/W>O#;1&',,+!'*$+,,5+$2!xLPK[[ et al.J!
CDD`yE!^#9!<+5,M5+&!10((2+!+5(!K95(0,A#$+'#,2'#,67!@09!VN,FH!"#![7$FH!eU,c/WC!3#9>
9+$5+$#()f! "45,67+(! U,c/WC! #(*! U,c/W]! '#,)+(#2"2! 4+$*+(J! #9! +5(+! O+&+125@52A2! *+,!
cX3W]!X(75;520$,!"#!+$$+567+(!xLPK[[ et al.J!CDD`n!Od3X et al.J!CDDGyE!!K(!*+$,+&;+(!L0,525>
0(! 85(*+2! ,567! "45,67+(! *+$!%2cPWC!#(*!*+$!-)c/WY!+;+(8'&&,! +5(!K95(0,A#$+'#,2'#,67!
DISKUSSION
H`!
@09!-&#FF! e%2cPWCf!"#!VN,FH! e-)c/WYfE!/'7+$! 5,2! '("#(+79+(J!*',,!*',,+&;+!P'250('&!
'#67! 8Q$!*5+!%(24561&#()!@0(! ,+&+125@+(!%2cPWC>X(75;520$+(! '()+4+(*+2!4+$*+(!1'((E!%,!
45$*! @+$9#2+2J! *',,! +5(! V'*#(),'#,2'#,67! "45,67+(! *+9! (+)'25@! )+&'*+(+(! -'9'2! #(*!
*+9!M0,525@!)+&'*+(+(!VN,5(!'*$+,,5+$2!4+$*+(!1'((J!#9!*5+!O+&+125@52A2!)+)+(!*5+!%2cPWC!
"#! +$7g7+(E! U5+$8Q$! ,5(*! 4+52+$+! dM2595+$#(),"N1&+(! (024+(*5)J! #9! +5(+(! ,+&+125@+(!
%2cPWC>X(75;520$!"#!+(24561+&(J!*+$!*5+,+(!V'*#(),'#,2'#,67!'*$+,,5+$2E!
SDU! "#%0.($0#:Q$'-.'+)3.!2%0!ZY*#(#.'2%3!0.'!".%1#(#0)1$9.!
?9!*5+!<+("595*'"0&>c'$;0(52$5&+!,2$#12#$;',5+$2!4+52+$!"#!0M2595+$+(J!4'$!+,!"#(A67,2!(02>
4+(*5)J!*+(!<5(*+90*#,!*5+,+$!O2$#12#$+(!'#8"#1&A$+(E!^#*+9!)5;2!*+$!<5(*+90*#,!456725>
)+!U5(4+5,+! *'$Q;+$J! 45+! *5+! K125@52A2! ,045+! *5+! O+&+125@52A2! 0*+$! '#67! *5+! %5)+(,67'82+(!
eK/B%!L$085&f!*5+,+$!X(75;520$+(!@+$;+,,+$2!4+$*+(!1g((2+(E!/#$67!Docking>K('&N,+(!10((>
2+!*+$!<5(*+90*#,!*+$!<+("595*'"0&>c'$;0(52$5&+!952!+5(+$!O#;,252#250(!'(!L0,5250(!PC!e,5+7+!
K;,67(522!YECE]f!(5672! +5(*+#25)! 5*+(2585"5+$2!4+$*+(E!U5+$;+5!4#$*+(!"4+5!9g)&567+!<5(*+>
90*5!5*+(2585"5+$2J!952!@+$)&+567;'$+(!Scoring>_+$2+(J!*5+!;+5*+!'&,!;50'125@+!W0(80$9'250(!
9g)&567! +$,675+(+(E! X9! *+9! +5(+(!<5(*+90*#,! ;5(*+(! *5+!<+("595*'"0&>c'$;0(52$5&+! Q;+$!
*+(!352$5&>O2561,2088!'(!*5+!Hinge>P+)50(!#(2+$!K#,;5&*#()!+5(+$!_',,+$,2088;$Q61+J!59!'(>
*+$+(!Q;+$!*+(!c'$;0(N&>O'#+$,2088E!%,!5,2!;+1'((2J!*',,!Scoring>:#(1250(+(!(#$!"#$!,67(+&>
&+(!<+4+$2#()!@0(!V5)'(*+(!5(!+5(+9!@5$2#+&&+(!Screening!*5+(+(!#(*!*'7+$!*5+!V5)'(*+(>
;5(*#()! (#$! )$0;! ';,67A2"+(E! O5+! ;+,52"+(! 9+5,2! (5672! '#,$+567+(*! (+)'25@+! <+52$A)+J! *5+!
*#$67! M0&'$+! 35672Q;+$+5(,2599#()+(! #(*! #(@0$2+5&7'82+! /+,0&@'2'250(! ,045+! '#67! *#$67!
,2+$5,67+!W0&&5,50(+(!"#,2'(*+!1099+(!x_KPP%3 et al.J!CDD`yE!?9!"#!#(2+$,#67+(J!45+!*+$!
4'7$,67+5(&567+! <5(*+90*#,! *+$! <+("595*'"0&>c'$;0(52$5&+! 5,2J! 4#$*+(! <+$+67(#()+(!
*#$67)+8Q7$2J!*5+!59!-+)+(,'2"!"#!Docking>K('&N,+(!'#67!*+(!+(2$0M5,67+(!<+52$')!*+$!<5(>
*#(),+(+$)5+!;+$Q61,56725)+(!x<KPPXV!k!OdVXRKJ!CDD`n!/%!KBdPXB et al.J!CDDFyE!^#$!
)+('#+(!?(2+$,#67#()!*+,!<5(*+@+$7'&2+(,!4#$*+(!:$+5+>%(+$)5+>O59#&'250(+(J!,M+"5+&&!*5+!
VX%>B+270*+! xKiRXO[!k!BKP%VX?OJ! CDDGy! +5()+,+2"2J! #9! '#67! *5+! (+)'25@+(!<+52$A)+!
+5(+$! V5)'(*+(;5(*#()! ,2A$1+$! "#! ;+$Q61,56725)+(! xKiRXO[! k!BKP%VX?OJ! CDDGn! 3%P>
RKVV et al.J!CDDFn!O[=%P3OcUK3[^ et al.J!CDD`yE!!
VX%>O59#&'250(+(!+$)';+(!8Q$!*$+5!R+$;5(*#()+(!*+$!<+("595*'"0&>c'$;0(52$5&+!+5(+!&+5672+!
<+@0$"#)#()! 8Q$! *+(! <5(*+90*#,J! 5(! *+9! *5+! V5)'(*+(! Q;+$! *5+! 352$5&)$#MM+! "#$!Hinge>
P+)50(!;5(*+(E!=+*067!5,2!*5+!/588+$+("!*+$!<5(*#(),+(+$)5+(!(5672!5(!+5(+$!-$gr+(0$*(#()J!
! ! DISKUSSION
HI!
*5+!+,!+$&'#;2J!,567+$!+5(+!d$5+(25+$#()!@0$7+$"#,')+(E!:Q$!"4+5!R+$;5(*#()+(!10((2+!1+5(+!
+5(*+#25)+!K#,,')+! )+2$088+(!4+$*+(J! *'! *5+!?(2+$,675+*+! "45,67+(! *+(!<5(*#(),+(+$)5+(!
*+$!;+5*+(!<5(*+90*5!5(!*+$!-$gr+(0$*(#()!*+,!O2'(*'$*8+7&+$,!&5+)+(E!%,!5,2!*+(1;'$J!*',,!
;+5*+!<5(*+90*5!9g)&567!,5(*E!%5(+!)+('#+!K#,,')+!Q;+$!*+(!<5(*+90*#,!*5+,+$!O2$#12#$+(!
*+$!<+("595*'"0&>c'$;0(52$5&+!1'((!,0952!(#$!+5(+!K#81&A$#()!*+$!W$5,2'&&,2$#12#$!*+,!L$02+>
5(>V5)'(*>W09M&+\+,!952!+5(+$!'#,$+567+(*!707+(!K#8&g,#()!&5+8+$(E!!!
/5+!4+52+$+!dM2595+$#()!*+$!<+("595*'"0&>c'$;0(52$5&+!&+)2+!*+(!<5(*+90*#,!8+,2E!K&&+!0M>
2595+$2+(! O2$#12#$+(! "+5)2+(! 59!Docking! +5(+! +5(7+52&567+! V5)'(*+(>L0,+E!/5+,+! O2$#12#$+(!
;5(*+(!5(!*+( Docking-%\M+$59+(2+(!Q;+$!*+(!c'$;0(N&>O2561,2088!Q;+$!+5(+!_',,+$,2088;$Q>
61+!"#$!Hinge>P+)50(E!
SDc! X$($9$3#.($0.99#.'2%3!0.'!B*IF=!2%0!0.'!>&IF=!
K#,)'(),M#(12! 8Q$! +5(! @5$2#+&&+,! Screening! ('67! X(75;520$+(!4'$+(!U090&0)5+90*+&&+! *+$!
%2BPW!#(*! *+$! L8BPWJ! *'! 1+5(+!W$5,2'&&,2$#12#$+(! *5+,+$! L$02+5(+! 5(! g88+(2&567+(!/'2+(>
;'(1+(!@+$8Q);'$!4'$+(E!%5(+!K('&N,+!*+$!BPW!L$02+5(+!@+$,675+*+(+$!d$)'(5,9+(!'#8!O+>
b#+("+;+(+!+$)';!+5(+(!(5+*$5)+(!;5,!9522&+$+(!W0(,+$@5+$#(),)$'*!e"45,67+(!YGq!#(*!YFq!
O+b#+("5*+(252A2fE!/5+!-Q2+!*+$!)+(+$5+$2+(!B0*+&&+!,0&&2+!952!*5+,+(!O+b#+("5*+(252A2+(!'#,>
$+567+(*!8Q$!+5(!90&+1#&'$+,!Docking!,+5(!xWPjOU[K:dRjcU et al.J!CDDIn!Bd?V[ et al.J!
CDDGyE!/5+!X*+(252A2,4+$2+!,5(*!"#*+9!'#8!*5+!-+,'92,+b#+("&A()+!;+"0)+(E!<+2$'672+2!9'(!
*5+!K[L><5(*+2',67+!,+M'$'2J! 5,2!*5+!W0(,+$@5+$#()!*+#2&567!7g7+$!e6'E!`Dq!O+b#+("5*+(25>
2A2fE!^#*+9!&5+)+(!'#67!*5+!O+b#+("5*+(252A2+(!*+$!,2$#12#$+&&!10(,+$@5+$2+(!P+)50(+(!eOcP,!
{!Structurally Conserved Regionsf!4+52!7g7+$!e6'E!FGqfE!!!
:Q$! +5(! @5$2#+&&+,!Screening!952! *+9!Docking>K(,'2"! 5,2! +,!456725)J! ;+5! (5672! @0$7'(*+(+$!
W$5,2'&&,2$#12#$! +5(! )#2+,! U090&0)5+90*+&&! '#8! <',5,! +5(+$! )++5)(+2+(! [+9M&'2,2$#12#$! "#!
)+(+$5+$+(E!K&,
DISKUSSION
HF!
/5+!)++5)(+2+(!W0(80$9'250(+(!*+$!K95(0,A#$+(!4+$*+(!,0952!@09![+9M&'2!'#8!*',!U090>
&0)5+90*+&&!Q;+$2$')+(E!!
/5+! U090&0)5+90*+&&5+$#()! +$80&)2+! #(2+$! 3#2"#()! +5(+$! 7#9'(+(! c/WC>[+9M&'2,2$#12#$!
eL/<>X/z!GdXPJ!xK3/%POd3 et al.J!CDDYyfE!%5(+!4+52+$+!5(!*+$!L/<>/'2+(;'(1!@+$"+567>
(+2+!7#9'(+!c/WI>W$5,2'&&,2$#12#$!eL/<>K66E>3$Ez!G?KCf!4#$*+!(5672!'&,![+9M&'2,2$#12#$!
8Q$!*5+!U090&0)5+90*+&&;5&*#()!+5()+,+2"2E!/5+!U,c/WI!;+,52"2!"4'$!+5(+!7g7+$+!O+b#+(">
5*+(252A2! "#! L8BPW!#(*!%2BPWJ! 5,2! ';+$! '#8)$#(*! *+$! )+$5()+$+(!K#8&g,#()! eY!hf!4+(5)!
)++5)(+2E!!
:Q$!*5+!U090&0)5+90*+&&5+$#()!4#$*+!952!*+$!O2$#12#$!GdXP!+5(+!5('125@+!:0$9!*+$!W5(',+!
@+$4+(*+2E! ^45,67+(! '125@+$! #(*! 5('125@+$! :0$9! )5;2! +,! )$0r+! ,2$#12#$+&&+! ?(2+$,675+*+E!
30;&+!et alE!)+7+(!*'@0(!'#,J!*',,!*5+!,2$#12#$+&&+!W0(,+$@5+$#()!;+5!*+$!5('125@+(!:0$9!)+>
$5()+$!5,2!'&,!;+5!*+$!'125@+(!:0$9!x3d<V% et al.J!CDD]yE!O0!1'((!;+5,M5+&,4+5,+!*5+!5('125@+!
:0$9!+5(+,!%("N9,!*#$67!+5(+(!'&&0,2+$5,67+(!B+67'(5,9#,!,2';5&5,+$2!4+$*+(E!/5+,!+$&+567>
2+$2! *5+!%(24561&#()! +5(+,! ,+&+125@+(! X(75;520$,J! 5(*+9!*5+,+$! +5(+! '&&0,2+$5,67+!P+)50(!*+$!
<5(*+2',67+!'*$+,,5+$2E!%5(!<+5,M5+&!75+$8Q$!5,2!*+$!_5$1,2088!X9'25(5;!e-&++@+6f!x/P?W%PJ!
CDDCyJ!*+$!+5(+!W0(80$9'250(,A(*+$#()!*+$!K;&!W5(',+!xKjd?< et al.J!CDDZy!;+45$12!#(*!
*'*#$67!*+(!5('125@+(!^#,2'(*!,2';5&5,5+$2!x3d<V% et al.J!CDD]yE!/5+,+!W0(80$9'250(,A(*+>
$#()!45$*!;+5! '(*+$+(!W5(',+(!(5672!7+$;+5)+8Q7$2! xW?<X3jX!k!BsVV%PJ!CDD]yE! =+*067!
1g((+(!'#67!)+$5()+$+!W0(80$9'250(,A(*+$#()+(!*+$!<5(*+2',67+!+5(+(!+(+$)+25,67!)Q(,25>
)+(!<+52$')!"#$!V5)'(*;5(*#()!;+5,2+#+$(E!U5+$*#$67!4A$+!+5(+!7g7+$+!O+&+125@52A2!)+)+(Q;+$!
'(*+$+(!W5(',+(!9g)&567!x=dU3Od3J!CDDHyE!/'*#$67!;+,52"2!*+$!_5$1,2088!+5(!)Q(,25)+$+,!
20\510&0)5,67+,!L$085&!'&,!'(*+$+J!;$+52!45$1,'9+!L$02+5(15(',+>X(75;520$+(!x:K<XK3!CDDZyE!!!
/5+!-Q2+!*+,!L8BPW!U090&0)5+90*+&&+,!10((2+!9522+&,!Docking Q;+$M$Q82!4+$*+(E!U5+$"#!
4#$*+(!;+1'((2+!L8BPW!d\5(*0&>X(75;520$+(!952!*+$!Docking>O0824'$+!-dV/!5(!*5+!K[L>
<5(*+2',67+!+5()+M',,2E!U5+$;+5!4#$*+(!M&'#,5;&+!V5)'(*+(>L0,+(!)+(+$5+$2J!4',!*5+!-Q2+!*+,!
)+(+$5+$2+(!U090&0)5+90*+&&,!;+,2A25)2E!!
SDR! Q)9#0#.'2%3!0.'!G$&*E)'.!>VW?MG!!
%5(!-$#(*!8Q$!*5+!K#,4'7&!@0(!LVK3[O!4'$!*5+!%885"5+("!*5+,+$!O0824'$+E!V52+$'2#$;+5,M5+>
&+!;+&+)+(J!*',,!*+$! 59M&+9+(25+$2+!Docking>K&)0$5279#,! 5(!*+$!V')+! 5,2J! +5(+!O#;,2'("*'>
2+(;'(1! ,+7$! ,67(+&&! ('67! M02+(25+&&+(! O#;,2'("+(! "#! *#$67,#67+(! xWdP< et al.J! CDD`n!
! ! DISKUSSION
HH!
WdP< et al.J!CDDHyE!%5(+!R'&5*5+$#()!*+$!O0824'$+!LVK3[O!4'$!'#8)$#(*!*+$!R+$8Q);'$1+52!
(#$!4+(5)+$!V52+$'2#$;+5,M5+&+!"#!@5$2#+&&+(!Screens!(024+(*5)E!!
^#$!B+270*+(@'&5*5+$#()! *+,! LVK3[O>Dockings! 4#$*+(! @+$,675+*+(+! R+$,#67+! *#$67)+>
8Q7$2J! *5+!*5+!-Q2+!*+,!@+$4+(*+2+(!L$0)$'99,!Q;+$M$Q8+(! ,0&&2+(E!K&,!P+8+$+("M$0)$'99!
*5+(2+! *5+!Docking>O0824'$+! -dV/J! *5+! ,670(! 082! 75(,5672&567! 57$+$!Performance! )+2+,2+2!
4#$*+!#(*!75+$;+5!)#2+!%$)+;(5,,+!+$"5+&2+! xd3d/%PK et al.J!CDDIn!Od?OK et al.J!CDD`yE!
K(7'(*!@0(!Re-Docking!%\M+$59+(2+(!4#$*+!Q;+$M$Q82J!0;!*5+!O0824'$+!LVK3[O!+;+(8'&&,!
)#2+! Docking>L0,+(! )+(+$5+$+(! 10((2+J! *E! 7E! *5+! +\M+$59+(2+&&! +$9522+&2+! L$02+5(>V5)'(*>
W0(80$9'250(! $+M$0*#"5+$2E! U5+$;+5! "+5)2+! ,567J! *',,! +,! ,0407&! 8Q$! LVK3[O! '&,! '#67! 8Q$!
-dV/!9g)&567!4'$J!8Q$!*5+!9+5,2+(!*+$!GZ!W$5,2'&&,2$#12#$+(J!*5+!V5)'(*+(>L0,+(!952!+5(+9!
PBO/>_+$2! @0(! Ç! C! h! "#! +$9522+&(J! 0;407&!Docking>L'$'9+2+$! @+$4+(*+2! 4#$*+(J! *5+!
,67(+&&+! <+$+67(#()+(! +$&'#;+(E! :Q$! +5(! @5$2#+&&+,! ScreeningJ! 5(! *+9! )$0r+! /'2+(;'(1+(!
('67!@5$2#+&&+(!Hits!*#$67,#672!4+$*+(J!5,2!+5(!95(59'&+$!P+67+('#84'(*!+5(!+(2,67+5*+(*+$!
:'120$E!/+$!5(!!LVK3[O!@+$4+(*+2+!K&)0$5279#,!+$&'#;2!+5(!+885"5+(2+$+,!Docking!#(*!+5(+!
)+('#+$+!L0,+(><+4+$2#()!'&,!-dV/E!/5+,!4#$*+!+;+(8'&&,!@0(!'(*+$+(!K#20$+(!;+,67$5+>
;+(!xWdP< et al.J!CDD`yE!!
X(! "4+5! *+$! YD! Re-Docking>%\M+$59+(2+! 10((2+! *5+! L$02+5(>V5)'(*+(>W0(80$9'250(! (5672!
$+M$0*#"5+$2!4+$*+(E!:Q$!*',!Re-Docking!5(!*5+!W$5,2'&&,2$#12#$!GUDG!4'$!LVK3[O!(5672!5(!
*+$!V')+J!+5(+!10$$+12+!L0,+!"#!)+(+$5+$+(E!/5+!O0824'$+!-dV/!$+M$0*#"5+$2+!8Q$!*+(!V5)'(>
*+(! *+$! W$5,2'&&,2$#12#$! GdXP! 1+5(+! 10$$+12+! V5)'(*+(>L0,+E! %5(! 9g)&567+$! -$#(*! *'8Q$!
1g((2+(!:+7&+$!5(!*+(!L$02+5(,2$#12#$+(!,+5(!xOd[PX::%P!k!/PKB<?P-J!CDDZyE!!
X(!('6780&)+(*+(!%\M+$59+(2+(!4#$*+(!*5+! 5(2+$(+(!<+4+$2#(),8#(1250(+(!LVK3[O>Score!
#(*!-dV/>Score!'#8!*+$+(!K(4+(*;'$1+52!8Q$!*5+!10$$+12+!<+4+$2#()!@0(!V5)'(*+(>L0,+(!
#(2+$,#672E!/5+!<+4+$2#()!*+$!Docking>Vg,#()+(!'(7'(*!*+,!;+$+67(+2+(!Scores!+$)';!+5(+!
)#2+! W0$$+&'250(! "45,67+(! -dV/>Ocore! #(*! *+9! +$$+67(+2+(! PBO/>_+$2E! /5+! Scoring>
:#(1250(!LVK3[O>Score!10$$+&5+$2+!p+*067!*+#2&567!;+,,+$!952!*+(!PBO/>_+$2+(!#(*!10((>
2+! ;+,,+$! "45,67+(! 10$$+12+(! #(*! 8'&,67+(! V5)'(*+(>L0,+(! *5,1$595(5+$+(! '&,! *5+! Scoring>
:#(1250(!-dV/>ScoreE!/5+,+!/5,1$595(5+$#()! 5,2! 8Q$!+5(!@5$2#+&&+,!Screening +(2,67+5*+(*J!
*'!75+$!082!(#$!*5+!;+,2;+4+$2+2+!L0,+!4+52+$!;+2$'672+2!45$*!xcPdOO et al.J!CDDHyE!!!
<+5!*+$!?(2+$,#67#()!*+,!85('&+(!Rankings!@0(!'125@+(!"#!5('125@+(!R+$;5(*#()+(J!4'$!*5+!
Scoring>:#(1250(! LVK3[O>Score! '#67! ;+,,+$! 5(! *+$! V')+! "45,67+(! '125@+(! #(*! 5('125@+(!
DISKUSSION
GDD!
R+$;5(*#()+(!"#!*5,1$595(5+$+(!'&,!*5+!Scoring>:#(1250(!-dV/>ScoreE!K&&+!*5+,+!%$)+;(5,>
,+!8Q7$2+(!"#!*+9!O67&#,,J!*',,!*5+!LVK3[O!O0824'$+!8Q$!+5(!@5$2#+&&+,!Screening!('67!X(75>
;520$+(! ^N1&5(>';7A()5)+$! W5(',+(! @'&5*+! 5,2E! /5+! Performance! *+$! O0824'$+! -dV/! #(*!
LVK3[O!1'((!8Q$!'(*+$+!L$02+5(1&',,+(!@'$55+$+(E!
SDe! Q#'*2.99.+!Screening!%),-!B*IF=:!2%0!>&IF=:/%-#;#*$'.%!!
3'67!s;+$M$Q8#()!*+$!%5)(#()!@0(!c/W>A7(&567+(!L$02+5(+(!'&,!^5+&M$02+5(+!"#$!%(24561>
&#()!(+#+$!K(25M'$',5251'J!4#$*+!952!U5&8+!67+905(80$9'25,67+$!B+270*+(!+5(+!O#67+!('67!
M02+(25+&&+(!X(75;520$+(!5(!+5(+$!)$0r+(!O#;,2'(";5;&5027+1!*#$67)+8Q7$2E!:Q$!*5+!O#67+!('67!
(+#+(!V+52,2$#12#$+(!4#$*+!*',!@5$2#+&&+!Screening!9522+&,!90&+1#&'$+(!Dockings!'&,!B+270>
*+! +5()+,+2"2J! 4+&67+! 7A#85)! K(4+(*#()! 85(*+2! x<ÑUB! k! OcU3%X/%PJ! CDDDn! cKRK>
Od[[d!k!dPPjJ!CDDIn!BcX33%OJ!CDDIn!d%VVX%3 et al.J!CDDHyE!:Q$!*',!@5$2#+&&+!Scree-
ning!('67!M02+(25+&&+(!%2BPW>!#(*!L8BPW>X(75;520$+(!4#$*+(!O2$#12#$,#67+(!5(!+5(+$!O#;>
,2'("*'2+(;'(1!*#$67)+8Q7$2J!*5+!R+$;5(*#()+(!+(27A&2J!4+&67+!;+$+52,!+5(+!K125@52A2!'#8!@+$>
,675+*+(+!W5(',+(!)+"+5)2!7';+(!xcU%3 et al.J!CDDGn!VX? et al.J!CDDIyE!/5+!K#,4'7&!*5+,+$!
W5(',+>801#,,5+$2+(! <5;&5027+1! +$7g72! *5+! c7'(6+J! %2BPW>! ,045+! L8BPW>X(75;520$+(! "#!
5*+(2585"5+$+(E!K#8)$#(*!*+$!,+7$!A7(&567+(!K[L><5(*+2',67+(!*+$!%2BPW!#(*!*+$!L8BPWJ!
4#$*+(!*$+5!O2$#12#$+(!59!@5$2#+&&+(!Screening! 5*+(2585"5+$2J!*5+!M$5("5M5+&&! 5(!*+$!V')+!,5(*!
;+5*+! L$02+5(+! "#! 5(75;5+$+(E! ^#*+9! 5,2! '("#(+79+(J! *',,! *5+! d\5(*0&>;',5+$2+(! L8BPW>
X(75;520$+(! '#67! *5+! %2BPW! 5(75;5+$+(! 1g((2+(J! *'! '#67! "4+5! *+$! 5*+(2585"5+$2+(! %2cPWC!
V+52,2$#12#$+(!'#8!*',!7090&0)+!L$02+5(!L8LWZ!+5(+!K125@52A2!"+5)2+(!e,5+7+!K;,67(522!YECEYfE!!!
SDHO! W2+;9#,5!
/5+!O+b#+(",#67+!('67!c/W!,045+!^N1&5(!7090&0)+(!L$02+5(+(!@0(!E. tenella!+$)';!9+7$+>
$+!(+#+!c/W,!,045+!^N1&5(+E!X9!4+52+$+(!L$0p+12@+$&'#8!4A$+!+,!4Q(,67+(,4+$2!'&&+!5*+(25>
85"5+$2+(!L$02+5(+!"#!1&0(5+$+(!#(*!"#!+\M$595+$+(E!%5(!4+52+$+,!K(,67&#,,+\M+$59+(2!4A$+!
*5+!/#$678Q7$#()!@0(!K125@5+$#(),>Assays! *+$!cPW,!952! *+(! ;+1'((2+(!^N1&5(+(J! #9!*5+!
p+4+5&5)+(!K125@5+$#(),M'$2(+$!*+$!cPW,!@0(!E. tenella! "#! 5*+(2585"5+$+(E!%5(!4+52+$+,!^5+&!
4A$+! +5(+! #98',,+(*+! ;5067+95,67+! c7'$'12+$5,5+$#()! *+$! c/W>! ,045+! ^N1&5(>A7(&567+(!
L$02+5(+! 5(! E. tenellaJ! #9! ;+5,M5+&,4+5,+! *5+! O#;,2$'2+! *+$! @+$,675+*+(+(! c/W>^N1&5(>
W09M&+\+!"#!5*+(2585"5+$+(E!/'$'#,!&5+r+!,567!9g)&567+$4+5,+!'#8!*+$+(!:#(1250(!59!L'$',5>
! ! DISKUSSION
GDG!
2+(!,67&5+r+(E!/'$Q;+$!75('#,!1g((2+!*5+!X*+(2585"5+$#()!4+52+$+$J!4+(5)+$!@+$4'(*2+$!c/W,!
@0(!E. tenella!9522+&,!90&+1#&'$;50&0)5,67+$!B+270*+(!+$80&)+(E!!
%5(+!@5$2#+&&+!V+52,2$#12#$0M2595+$#()!*+$!<+("595*'"0&>c'$;0(52$5&+!4#$*+!;+$+52,!*#$67)+>
8Q7$2E! X9! 4+52+$+(! L$0p+12@+$&'#8! ,0&&2+! *5+! 67+95,67+!Optimierung! 8Q$! '&&+! 5*+(2585"5+$2+(!
V+52,2$#12#$+(!e<+("595*'"0&+J!<+("595*'"0&+>c'$;0(52$5&+J!P70*'(5(>d\5(*0&+!#(*!3'M720>
&'62'9+f!*#$67)+8Q7$2!4+$*+(J!,045+!*5+,+!0M2595+$2+(!V+52,2$#12#$+(!45+*+$#9!+\M+$59+(>
2+&&J!*',!7+5r2!in vitro!'9!^5+&M$02+5(!)+2+,2+2!4+$*+(E!B52!U5&8+!*+$!,0!)+40((+(+(!X(80$9'>
250(+(!Q;+$!*5+!K125@52A2!*+$! +5("+&(+(!O2$#12#$+(J!1g((2+(!*'((!*5+!L7'$9'10M70$90*+&&+!
,045+!*5+!Docking>L'$'9+2+$!@+$8+5(+$2!4+$*+(!,045+!'#67!*5+!VX%>L'$'9+2+$!'()+M',,2!4+$>
*+(E!!!
UNM027+,+(!Q;+$!9g)&567+!<5(*#(),90*5!*+$!V5)'(*+(!5(!*+$!K[L><5(*+2',67+!1g((+(!952!
U5&8+!@0(!U090&0)5+90*+&&+(!9+5,2!(#$!;+*5()2!';)+&+52+2!4+$*+(!xcKRKOd[[d!k!LUK>
[KWJ!CDDHyE!K#,!*5+,+9!-$#(*!,5(*!+5(+!;"4E!@+$,675+*+(+!%2cPWC>X(75;520$109M&+\+!"#!
+9M8+7&+(E!^#*+9!4A$+(!'#67!O2$#12#$'#81&A$#()+(!*+$!7#9'(+(!c/WC!952!*+(!5*+(2585"5+$>
2+(! V+52,2$#12#$+(! '&,! 60>1$5,2'&&5,5+$2+! V5)'(*+(! 4Q(,67+(,4+$2E! /5+,+! %$)+;(5,,+! ,0&&2+(!
456725)+!U5(4+5,+!8Q$!*5+!+\M+$59+(2+&&+!R0$)+7+(,4+5,+!&5+8+$(E!!
/'$Q;+$!75('#,!1g((2+!+5(+!67+95,67+!dM2595+$#()!*+$!+$9522+&2+(!V+52,2$#12#$+(!+$,2+$!-+>
(+$'250(!#(2+$!<+$Q61,56725)#()!*+$!L'$'9+2+$
DISKUSSION
GDC!
;+$+52,!*#$67)+8Q7$2E!:#(1250(,,2#*5+(J!45+!"E<E!2$'(,)+(+!s;+$+\M$+,50(!0*+$!knock out!B#>
2')+(+,+! xV%_X3J! CDDFy! '(! E. tenella! ,0&&2+(! *'$Q;+$! 75('#,! 1&A$+(J! 4+&67+! P0&&+! *5+!
%2cPWC!,045+!*5+!%2BPW!59!^+&&"N1&#,!@0(!E. tenella!,M5+&2E!!
! ! ZUSAMMENFASSUNG
GDY!
T!72+)((.%&)++2%3!L'$',52+(!*+$!Apicomplexa!#98',,+(!,0407&!9+7$+$+!7#9'(M'270)+(+J!'&,!'#67!25+$M'270)+>
(+! L$020"0+(E! <+5,M5+&+! 8Q$! 456725)+! R+$2$+2+$! 7#9'(>! #(*! 25+$M'270)+(+$! L'$',52+(! ,5(*!
Plasmodium falciparum!#(*!Eimeria tenellaE!/#$67!P. falciparum!45$*!*5+!2$0M5,67+!X(8+125>
0(,1$'(17+52!Malaria tropica!7+$@0$)+$#8+(J!*5+!pA7$&567!6'E!CJI!B5&&50(+(![0*+,0M8+$!80$*+$2!
xB?PLUj!k!<P%BK3J!CDDGyE!E. tenella!@+$#$,'672!W01"5*50,+! 5(!UQ7(+$(J!*5+!4+&24+52!
8Q$!R+$&#,2+!5(!+5(+$!)+,67A2"2+(!Ug7+!"45,67+(!ZDD!B5&&50(+(!u!xOUXPV%j et al.J!CDDIy!#(*!
#(8',,;'$+(! Y! B5&&5'$*+(! ?Ov! x/KVVd?V! k! VXVV%Ud=J! CDD`y! @+$'(240$2&567! "+567(+2E!
%5(+! M$0M7N&'125,67+! R'1"5(5+$#()! )+)+(! *5+,+! W$'(17+52! 5,2! 9+5,2! g10(095,67! 5(+88+125@!
xWX33KXP/ et al.J!CDD]n!Bc/d3KV/!k!OUXPV%jJ!CDDHyJ!#(*!+5(+!<+7'(*&#()!952!W01>
"5*50,2'251'! 45$*! *#$67! 7A#85)+! P+,5,2+(";5&*#()! )+)+(! ;+1'((2+! _5$1,2088+! +$,674+$2!
xWX33KXP/ et al.J!CDD]n!B%UVUdP3 et al.J!GHHYn!B%V%3/%^J!CDDDn!OUXPV%jJ!GHFHyE!
/5+,! +$80$*+$2! *5+! %(24561&#()! (+#+$! M$+5,4+$2+$! K&2+$('25@+(E! -++5)(+2+! ^5+&M$02+5(+! "#$!
%(24561&#()! (+#'$25)+$!K$"(+5,2088+! ,5(*! *5+!^N1&5(>';7A()5)+(!W5(',+(! ec/WfJ! "#! *+(+(!
'#67!*5+!c/W>related!W5(',+!C!e%2cPWCf!'#,!E. tenella!)+7g$2E!/5+,+!L$02+5(+!,5(*!9'r)+;>
&567!'(!*+$!P+)#&'250(!*+,!^+&&"N1&#,!;+2+5&5)2! x3%VOd3!k!cd}J!CDDGyE!/#$67!67+95,67+!
R'&5*5+$#()! 952! *+9! c/W>X(75;520$! :&'@0M5$5*0&! 10((2+! ('67)+45+,+(! 4+$*+(J! *',,! +5(!
:#(1250(,@+$&#,2! @0(!c/W,! 5(!E. tenella! *5+!R+$9+7$#()!*+,!L'$',52+(! 5(!W#&2#$! 5(75;5+$2E!
c/W,!5(!E. tenella!,5(*!*'7+$!'&,!^5+&M$02+5(+!8Q$!*5+!%(24561&#()!+5(+$!c7+9027+$'M5+!*+$!
W01"5*50,+!)++5)(+2E!!!
/5+! 5(! *5+,+$!K$;+52! *#$67)+8Q7$2+(! ;505(80$9'25,67+(![5+8+('('&N,+(! ,0&&2+(! *',!R0$109>
9+(!@0(!c/W!L$02+5(+(!59!L'$',52+(!E. tenella!Q;+$M$Q8+(E!/',!-+(09!@0(!E. tenella!&5+)2!
'&,!+5(!+$,2+$!%(24#$8!@0$!eDraft Assemblyf!x82Mzoo82ME,'()+$E'6E#1yE!/5+!%$,2+&&#()!*+$![5+8+(>
'('&N,+! *+$! 7090&0)+(! c/W! L$02+5(+! +$80&)2+! *#$67! O'99&#()! #(*! <+4+$2#()! @0(! O+>
b#+("5(80$9'250(+(!@0(!'(*+$+(!d$)'(5,9+(!*+,!O2'99+,!*+$!ApicomplexaE!/'"#!4#$*+(J!
(+;+(! *+$!K;8$')+! ;50&0)5,67+$!/'2+(;'(1+(! ,045+! V52+$'2#$*'2+(;'(1+(J! '#67! +5(+!P+57+!
4+52+$+$! ;505(80$9'25,67+$! B+270*+(J! 45+! M''$4+5,+! #(*! 9#&25M&+! O+b#+("@+$)&+567+! eA-
lignmentsfJ! '()+4'(*2! x[UdBLOd3 et al.J!GHH]yE!/'$Q;+$!75('#,!4#$*+!+5(+!K('&N,+!*+$!
+(2,M$+67+(*+(!^N1&5(+!>!*+$!K125@'20$+(!*+$!c/W,!>!;+"Q)&567!:#(1250(!#(*!O2$#12#$J!,045+!
+5(+! /'2+(;'(1,#67+! ('67! ;5,7+$! (5672! ;+,67$5+;+(+(! ^N1&5(+(! 5(!E. tenella! *#$67)+8Q7$2E!
/#$67!*5+,+!;505(80$9'25,67+(!K('&N,+(!10((2+(!(+;+(!*+$!;+$+52,!;+1'((2+(!%2cPWCJ!*$+5!
4+52+$+J!;5,&'()!(5672!'((025+$2+!c/W,!5(!E. tenella! 5*+(2585"5+$2!4+$*+(!e%2cPWGJ!%2cPWY!
ZUSAMMENFASSUNG
GD]!
,045+!%2BPWfE!_+52+$+!<VKO[>O#67+(!+$)';+(!@5+$!(+#+!M02+(25+&&+!^N1&5(+!8Q$!E. tenellaJ!
40@0(!%2cjcY'!'&,!K125@'20$!*+$!%2cPWC!@0(!B'$�'!VE!O#Ä$+"!:+$(Ä(*+"!eX(2+$@+2!X((0@'>
250(!-9;UJ!O674';+(7+59f!;+,2A25)2!4+$*+(!10((2+E!<505(80$9'25,67+!K('&N,+(!&',,+(!@+$>
9#2+(J!*',,!'#67!%2cjcG!#(*!%2cjcY;!5(!*+$!V')+!,5(*!%2cPWC!"#!'125@5+$+(E!K#r+$*+9!
5,2!'("#(+79+(J!*',,!%2cjc]!'&,!K125@'20$!*+$!%2cPWG!8#()5+$2E!!
X(75;520$+(!*+$!%2cPWCJ!*5+!5(!@0$'()+)'()+(+(!K$;+52+(!)+8#(*+(!4#$*+(J!10((2+(!;+$+52,!
5(! +("N9'25,67+(
! ! LITERATURVERZEICHNIS
GDZ!
U!V#*.')*2'6.'1.#,-%#+!!
K66+&$N,!X(6EJ!OE!/E!?EE!X(,5)72!XXJ!B0*+&&5()!%(@5$0(9+(2E!!CDDYE!!
K&9&08J!BEJ!<$'(*,*'&J!<E!dEJ!'(*!Kb@5,2J!=EJ!CDD]J!<5(*5()!'885(52N!M$+*56250(!4527!*588+$+(2!80$6+!85+&*,z!+\'95('250(!08!27+!&5(+'$!5(2+$'6250(!+(+$)N!9+270*z!=!c09M#2E!c7+9J!JT\!GC]C>GCZ]E!
K&2,67#&J!OE!:EJ!-5,7J!_EJ!B5&&+$J!_EJ!BN+$,J!%E!_EJ!'(*!V5M9'(J!/E!=EJ!GHHDJ!<',56!&06'&!'&5)(9+(2!,+'$67!200&z!=!B0&!<50&J!JHT\!]DY>]GDE!
K(*+$,+(J!-EJ!<#,,0J!/EJ!L02+$,"9'(J!KEJ!U4'()J!=E!PEJ!_#$2"J!=E!BEJ!P5MMJ!PEJ![75+$$NJ!=E!cEJ!%)&NJ!=E!BEJ!'(*!B0$',J!/EJ!GHHIJ![7+!,2$#62#$+!08!6N6&5(!Uz!60990(!90*+!08!15(',+!'625@'250(!'(*!,M+65856!8+'2#$+,z!%B<d!=J!HU\!HZF>H`IE!
K(*+$,0(J!BEJ!<+'225+J!=E!:EJ!<$+'#&2J!-E!KEJ!<$++*J!=EJ!<N27J!WE!:EJ!c#&,7'4J!=E!/EJ!%&&,20(J!PE!LEJ!-$++(J!OEJ!B5(,7#&&J!cE!KEJ!30$9'(J!PE!KEJ!L'#M252J!PE!KEJ!O2'(4'NJ!=EJ![709',J!KE!LEJ!'(*!=+4,;#$NJ!LE!=EJ!CDDYJ!X95*'"0xGJC>'yMN$5*5(+,z!'!M02+(2!'(*!,+&+625@+!6&',,!08!6N6&5(>*+M+(*+(2!15(',+!5(75;520$,!5*+(2585+*!27$0#)7!,2$#62#$+>;',+*!7N;$5*5,'250(z!<500$)E!B+*!c7+9!V+22J!HP\!YDCG>YDC`E!
Kb@5,2J!=EJ!V#"710@J!RE!<EJ!'(*!<$'(*,*'&J!<E!dEJ!CDDCJ!V5)'(*!;5(*5()!'885(525+,!8$09!B/!,59#&'250(,z!K66E!c7+9!P+,J!PT\!YZF>Y`ZE!
Kb@5,2J!=E!'(*!B'$+&5#,J!=EJ!CDDGJ![7+!&5(+'$!5(2+$'6250(!+(+$)N!9+270*!80$!M$+*5625()!&5)'(*!;5(*5()!8$++!+(+$)5+,z!c09;!c7+9!U5)7![7$0#)7M#2!O6$++(EJ!S\!`GY>`C`E!
K$;5,+$J!=E!VE!'(*!K9;&'$*J!:EJ!CDD`J!U0(0150&!/+$5@'25@+,!80$!27+![$+'29+(2!08!L$0&58+$'25@+!/5,0$*+$,z!L[co?OCDD`oDD`]H]E!
K$@5+#\J!cE!'(*![$5;#2J!dEJ!CDDZJ!K9M$+('@5$!0$!80,'9M$+('@5$!M&#,!$520('@5$!5(!UXR!5(8+6>250(z!M7'$9'60&0)NJ!+8856'6N!'(*!20&+$';5&52N!M$085&+z!/$#),J!UT\!`YY>`ZHE!
KN0#;J!_E!OEJ!-+&&+$J!OE!KEJ![$'(J![EJ!B'$25(J!LEJ!R5+$&5()J!=E!BEJ!'(*!L00$*'*J!:E!:EJ!CDDZJ!X9'25(5;!e-&++@+6f>5(*#6+*!7+M'2020\5652Nz!=!c&5(!-',2$0+(2+$0&J!Pe\!IZ>IIE!
<'*'4+NJ!%E!KE!'(*!W'MM+J![EJ!GHHHJ!<+("595*'"0&+!60(*+(,+*!$5()!,N,2+9,E!}XE!ON(27+,5,!08!,09+!,#;,252#2+*!6N6&0'&1N&!MN$5*0xGJC>'y;+("595*'"0&+,!4527!'(2565M'2+*!'(25(+0>M&',256!'625@52Nz!%#$!=!B+*!c7+9J!PS\!``Y>``IE!
<'$+,675(0J!BE!KEJ!O67+225(0J!cEJ![$05'(5J![EJ!B'$25(+&&5J!%EJ!B0$)5&&0J!:EJ!'(*!c5'$*5+&&0J!:EJ!CDDIJ!%$&025(5;!5(!6'(6+$!2$+'29+(2z!K((!d(60&EJ!HR\!YZ>]GE!
<'$$5&J!}E!'(*!O0&5@'J!PEJ!CDD`J!B0&+6#&'$!90*+&&5()z!B0&!<50,N,2EJ!J\!``D>`FGE!
<',2'$$'67+'J!PE!KEJ!B0(2+$0J!=E!cEJ!O''@+*$'>-'p'$*0J!XEJ!c+$*'>:&0$+,J!PEJ!B'67'*0>/095()#+"J!KEJ!'(*!c09#""5+J!KE!-EJ!CDDFJ!xB0&+6#&'$!2'$)+2,!80$!(+4!*$#)!*5,60@+$N!20!2$+'2!2NM+!C!*5';+2+,!'(*!0;+,52Nyz!P+@!B+*!c75&EJ!HPU\!GDI>GGIE!
LITERATURVERZEICHNIS
GD`!
<'2+9'(J!KEJ!c05(J!VEJ!/#$;5(J!PEJ!:5((J!PE!/EJ!U0&&567J!REJ!-$588527,>=0(+,J!OEJ!W7'(('J!KEJ!B'$,7'&&J!BEJ!B0\0(J!OEJ!O0((7'99+$J!%E!VEJ!O2#*70&9+J!/E!=EJ!j+'2,J!cEJ!'(*!%**NJ!OE!PEJ!CDD]J![7+!L8'9!M$02+5(!8'95&5+,!*'2';',+z!3#6&+56!K65*,!P+,J!PJ\!/GYF>/G]GE!
<'N,J!UE!%EJ!CDD]J!c#$$+(2!'(*!5(@+,25)'250('&!'(250;+,52N!')+(2,!'(*!0;+,52N!27+$'M+#256!2$+'29+(2!2'$)+2,z!d;+,E!P+,J!HJ\!GGHI>GCGGE!
<+95,J!-E!_E!'(*!B#$610J!BE!KEJ!GHH`J![7+!M$0M+$25+,!08!1(04(!*$#),E!GE!B0&+6#&'$!8$'9+>40$1,z!=!B+*!c7+9J!Pe\!CFFI>CFHYE!
<+(,0(J!/EJ!V5M9'(J!/E!=EJ!'(*!d,2+&&J!=EJ!GHHYJ!-+(<'(1z!3#6&+56!K65*,!P+,J!JH\!CH`Y>CH`ZE!
<+$9'(J!UEJ!U+($561J!WEJ!'(*!3'1'9#$'J!UEJ!CDDYJ!K((0#(65()!27+!40$&*45*+!L$02+5(!/'2'!<'(1z!3'2!O2$#62E!<50&J!HO\!HFDE!
<+$27+2J!cE!'(*!W'&*5,J!LEJ!CDDIJ!c+&&>,M+65856!$+,M0(,+,!20!&0,,!08!6N6&5(>*+M+(*+(2!15(',+,z!d(60)+(+J!JU\!]]`H>]]IIE!
<+N+$J!cEJ!CDDIJ!%2';&5+$#()!+5(+$!-$5*>c09M#25()!?9)+;#()!#(*!@5$2#+&&+,!O6$++(5()!'(!cN6&5(>';7A()5)+(!W5(',+(!@0(!@+2+$5(A$>!#(*!7#9'(>$+&+@'(2+(!L'$',52+(z!/5,,+$250(!59!:'67;+$+567!c7+95+!#(*!L7'$9'"5+!*+$!%;+$7'$*>W'$&,>?(5@+$,52A2![Q;5()+(E!
<0+619'((J!<EJ!<'5$067J!KEJ!KM4+5&+$J!PEJ!<&'22+$J!BE!cEJ!%,2$+567+$J!KEJ!-',2+5)+$J!%EJ!B'$>25(J!BE!=EJ!B5670#*J!WEJ!dâ/0(0@'(J!cEJ!L7'(J!XEJ!L5&;0#2J!OEJ!'(*!O67(+5*+$J!BEJ!CDDYJ![7+!O_XOO>LPd[!M$02+5(!1(04&+*)+;',+!'(*!52,!,#MM&+9+(2![$%B<V!5(!CDDYz!3#6>&+56!K65*,!P+,J!PH\!Y`Z>YIDE!
<g79J!UE!=EJ!W&+;+J!-EJ!'(*!W#;5(N5J!UEJ!CDDCJ!_5$1,2088*+,5)(z!OM+12$#9!K1'*E!R+$&')J!U+5*+&;+$)J!<+$&5(J!d\80$*EJ!H\!]CI>]ZFE!
<g79J!UE!=E!'(*!O67(+5*+$J!-EJ!CDDDJ!R5$2#'&!O6$++(5()!80$!<50'625@+!B0&+6#&+,z!_5&+N>RcU!R+$&')J!_+5(7+59E!
<0#&*J!=E!-EJ!%&,7+517'J!UE!BEJ!'(*!B0$,NJ![E!KEJ!CDDHJ!K@5'(!60665*50,5,z!27+!;',56!M'270&>0)N!20!60(2$0&z!=!%)NM2!O06!L'$',520&J!Pe\!FZ>HFE!
<$'6675>P56'$*J!REJ!<'$51J!OEJ!/+&@+66750J!cEJ!/0+$5)J!cEJ!c7'1$';'$25J!PEJ!'(*!c7'1$';'$25J!/EJ!CDDDJ!L8LW`J!'!(0@+&!6N6&5(>*+M+(*+(2!15(',+o9520)+(>'625@'2+*!M$02+5(!15(',+>$+&'2+*!M$02+5(!15(',+!8$09!L&',90*5#9!8'&65M'$#9z!<5067+9!=J!PSc\!CZZ>C`YE!
<$'9,0(J!UE!3EJ!c0$0('J!=EJ!/'@5,J!OE![EJ!/561+$,0(J!OE!UEJ!%*+&,2+5(J!BEJ!:$N+J!OE!REJ!-'9M+J!PE![EJ!=$EJ!U'$$5,J!LE!KEJ!U',,+&&J!KEJ!U0&9+,J!_E!/EJ!U#(2+$J!PE!3EJ!V'61+NJ!WE!%EJ!V0@+p0NJ!<EJ!V#""50J!BE!=EJ!B0(2'('J!REJ!P06b#+J!_E!=EJ!P#,('1J!/EJ!O7+467#1J!VEJ!R+'&J!=E!BEJ!_'&1+$J!/E!UEJ!'(*!W#NM+$J!VE!:EJ!CDDGJ!d\5(*0&+>;',+*!5(75;520$,!08!6N>6&5(>*+M+(*+(2!15(',+!C!ec/WCfz!*+,5)(J!,N(27+,5,J!+("N9'256!'625@525+,J!'(*!}>$'N!6$N,2'&&0)$'M756!'('&N,5,z!=!B+*!c7+9J!SS\!]YYH>]YZFE!
<$'(*+(J!cE!'(*
! ! LITERATURVERZEICHNIS
GDI!
<$'@+J!BEJ!-00*9'(J!REJ!W'95(,1',J!%EJ!:'$$+&&J!KEJ![599+$J!_EJ!L0M+J!OEJ!U'$'M'(7'&&5J!PEJ!O';+$J!UEJ!B0$,+J!/EJ!<#&&061J!=EJ!B+(J!KEJ!300$NJ!cEJ!P'967'(*'(5J!PEJ!W+(('J!VEJ!<0027J!<EJ!-0;;#$#J!=EJ!=5'()J!}EJ!O$5*7'$'J!PEJ!=#,256+J!PEJ!'(*!L'"*#$J!PEJ!CDDFJ!OM$N>6+&!80$!67$0(56!9N+&05*!&+#1+95'!'(*!L75&'*+&M75'!67$090,09+>M0,525@+!'6#2+!&N9>M70;&',256!&+#1+95'!$+,5,2'(2!20!0$!5(20&+$'(2!08!59'25(5;!9+,N&'2+z!c&5(!c'(6+$!P+,J!HS\!YZC>YZHE!
<$+52+(&+67(+$J!cE!<EJ!<0,,+9+N+$J!/EJ!'(*!%()7J!PE!KEJ!CDDZJ!c$N,2'&&0)$'M7N!80$!M$02+5(!15(',+!*$#)!*+,5)(z!LWK!'(*!OPc!6',+!,2#*5+,z!<506759E!<50M7N,E!K62'J!HcTS\!YF>]HE!
<$5619'((J!=EJ!GHHZJ!X(2+$'625@+!@5,#'&5"'250(!08!90&+6#&'$!,6+('$50,!4527!BdV>cK/oOj<jVE!X(z!/'2'!@5,#'&5"'250(!5(!90&+6#&'$!,65+(6+z![00&,!80$!5(,5)72!'(*!5((0@'>250(Ez!K**5,0(>_+,&+N!L#;&E!c09MJ!P+'*5()J!B',,J!FY>HIE!
<$5619'((J!=EJ!W+5&J!BEJ!%\(+$J![E!%EJ!'(*!B'$7g8+$J!PE!=EJ!CDDDJ!B0&+6#&'$!-$'M756,z![$+(*,!'(*!L+$,M+625@+,Ez!=E!B0&E!B0*+&EJ!U\!YCF>Y]DE!
<$0M7NJ!/E!:EJ!X,$'+&J!/E!OEJ!L',20$J!KEJ!-5&&025(J!cEJ!c7522561J!-E!%EJ!ON90(*,J!_E![EJ!V0#J!jEJ!O'*&+$J!<E!BEJ!'(*!L0&1J!PE!%EJ!CDDDJ!L7'$9'6015(+256!5(2+$'6250(!;+24++(!'9M$+('@5$!'(*!6&'$527$09N65(!5(!7+'&27N!9'&+!@0&#(2++$,z!K(25956$0;!K)+(2,!c7+9027+$J!SS\!HIF>HF]E!
<#5p,9'(J!PEJ!CDD]J!c7+90)+(0956,!5(!/$#)!/5,60@+$Nz!O2$#62#$'&!K,M+62,!08!W5(',+,!'(*![7+5$!X(75;520$,z!_5&+N>RcU!R+$&')!-9;U!k!c0E!W-'KJ!JJ\!GHG>CG`E!
<#$)+J!cE!'(*!W'$&5(J!OEJ!GHHIJ!L$+*56250(!08!609M&+2+!)+(+!,2$#62#$+,!5(!7#9'(!)+(0956!/3Kz!=!B0&!<50&J!JUR\!IF>H]E!
c'5J!}EJ!:#&&+$J!KE!VEJ!B6/0#)'&*J!VE!PEJ!'(*!^7#J!-EJ!CDDYJ!KM560M&',2!)+(09+!08!27+!606>65*5'(!%59+$5'!2+(+&&'z!-+(+J!PJH\!YH>]`E!
c'(*#$5J!:EJ!L+$+"J!LE!cEJ!c'6+$+,J!PE!KEJ!'(*!*+J!KE!_EJ!=$EJ!CDDIJ!L$02+5(!15(',+,!',!2'$)+2,!80$!'(25M'$',5256!67+9027+$'MN!*$#),z!c#$$!/$#)!['$)+2,J!R\!YFH>YHFE!
c'$&,0(J!_EJ!W'$M&#,J!BEJ!'(*!U';+$J!%EJ!GHFZJ!c0(,2$#6250(!08!'!90*+&!80$!27+!27$++>*59+(,50('&!,2$#62#$+!08!7#9'(!$+('&!$+(5(z!UNM+$2+(,50(J!c\!GY>C`E!
c'@',0220J!cE!3EJ!W0@'6,J!=E!KEJ!'(*!K;')N'(J!PE!KEJ!CDDZJ!P+M$+,+(25()!$+6+M20$!8&+\5;5&52N!5(!&5)'(*!*0615()!27$0#)7!$+&+@'(2!(0$9'&!90*+,z!=!K9!c7+9!O06J!HJc\!H`YC>H`]DE!
c'@',0220J!cE!3E!'(*!d$$NJ!KE!=EJ!CDDIJ!V5)'(*!*0615()!'(*!,2$#62#$+>;',+*!@5$2#'&!,6$++(5()!5(!*$#)!*5,60@+$Nz!c#$$![0M!B+*!c7+9J!c\!GDD`>GDG]E!
c'@',0220J!cE!3E!'(*!L7'2'1J!OE!OEJ!CDDHJ!U090&0)N!90*+&5()!5(!*$#)!*5,60@+$Nz!6#$$+(2!2$+(*,!'(*!'MM&56'250(,z!/$#)!/5,60@
LITERATURVERZEICHNIS
GDF!
c7+(J!jEJ!=5$')+J!/EJ!c'$5*7'J!/EJ!W'276'$2J!KE!WEJ!c0$2+,J!%E!KEJ!/+((#&&J!PE!KEJ!-+N+$J!=E!KEJ!L$5))+J!OE![EJ!'(*!_'2+$,J!3E!cEJ!CDD`J!X*+(25856'250(!08!'(!+88+620$!M$02+5(!'(*!)'5(>08>8#(6250(!9#2'(2,!27'2!'625@'2+!L89$1J!'!9'&'$5'&!6N6&5(>*+M+(*+(2!M$02+5(!15(',+z!B0&!<5067+9!L'$',520&J!HSe\!]F>ZIE!
c70()!_+,&+NJ!WE!BEJ!c7#J!OE!OEJ!V5J!VEJ!/#@'*5+J!PE!WEJ!j'()J!jEJ!'(*!}5'0J!_EJ!CDDYJ!O#;>,252#2+*!]>'95(0>275'"0&>C>N&!609M0#(*,!',!6N6&5(>*+M+(*+(2!15(',+!5(75;520$,z!?O!L'2>+(2!`Z`HFIFE!
c70#7'(J!=E!/EJ!^'9'$$5M'J!/E!%EJ!V'5J!LE!UEJ!d$'9',50(4#J!cE!?EJ!'(*!-$';5(,15J!=E!VEJ!CDDIJ!O#(525(5;!eO#2+(2fz!'!(0@+&!')+(2!80$!27+!2$+'29+(2!08!9+2',2'256!$+('&!6+&&!6'$65(09'z!=!d(60&E!L7'$9!L$'62J!HP\!Z>GZE!
c&'$1J!PE!/EJ!CDDHJ!L$0,M+625@+!&5)'(*>!'(*!2'$)+2>;',+*!Y/!iOKPz!,2'2+!08!27+!'$2!CDDFz!c#$$![0M!B+*!c7+9J!e\!IHG>FGDE!
c0)0&&#*0J!KEJ!L+$+">R5"6'5(0J!:EJ!'(*!['9'$)0J!=EJ!CDDZJ!3+4!5(,5)72,!5(!27+!M7'$9'60&0)5>6'&!27+$'MN!08!'$2+$5'&!7NM+$2+(,50(z!c#$$!dM5(!3+M7$0&!UNM+$2+(,EJ!HS\!]CY>]CIE!
c07+(J!:E!%E!'(*!O2+$(;+$)J!BE!=EJ!GHFDJ!d(!27+!M$+*56250(!08!M$02+5(!,2$#62#$+z![7+!,5)(585>6'(6+!08!27+!$002>9+'(>,b#'$+!*+@5'250(z!=!B0&!<50&J!HPR\!YCG>YYYE!
c$0,,J!=E!<EJ![709M,0(J!/E!cEJ!P'5J!<E!WEJ!<';+$J!=E!cEJ!:'(J!WE!jEJ!U#J!jEJ!'(*!U#9;&+2J!cEJ!CDDHJ!c09M'$5,0(!08!,+@+$'&!90&+6#&'$!*0615()!M$0)$'9,z!M0,+!M$+*56250(!'(*!@5$2#'&!,6$++(5()!'66#$'6Nz!=!c7+9!X(8!B0*+&J!Se\!G]ZZ>G]I]E!
c#$$NJ!UE!<EJ!GH]]J![7+!9+270*!08!,2++M+,2!*+,6+(2!80$!(0(&5(+'$!95(595"'250(M$0;&+9,Ez!i#'$2E!KMM&E!B'27EJ!J\!CZF>C`YE!
/'&&0#&J!PE!KE!'(*!V5&&+70pJ!UE!OEJ!CDD`J!L0#&2$N!60665*50,5,z!$+6+(2!'*@'(6+9+(2,!5(!60(2$0&!9+',#$+,!'(*!@'665(+!*+@+&0M9+(2z!%\M+$2!P+@!R'665(+,EJ!T\!G]Y>G`YE!
/'@5+,J![E!-EJ![#(('7J!LEJ!B+5p+$J!VEJ!B'$10J!/EJ!%5,+(;$'(*J!-EJ!%(*56022J!=E!KEJ!'(*!30;&+J!BE!%EJ!CDDGJ!X(75;520$!;5(*5()!20!'625@+!'(*!5('625@+!c/WCz!27+!6$N,2'&!,2$#62#$+!08!c/WC>6N6&5(!Ko5(*5$#;5(>Z>,#&M70('2+z!O2$#62#$+J!e\!YFH>YHIE!
*+!K90$59J!UE!VEJ!c'6+$+,J!PE!KEJ!'(*!3+2"J!LE!KEJ!CDDFJ!V5(+'$!5(2+$'6250(!+(+$)N!eVX%f!9+270*!5(!&+'*!*5,60@+$N!'(*!0M2595"'250(z!c#$$!/$#)!['$)+2,J!e\!GGDD>GGDZE!
/+'(J!LE!BEJ!GHH]J!P+6+(2!'*@'(6+,!5(!*$#)!*+,5)(!9+270*,z!47+$+!45&&!27+N!&+'*éz!<50+,>,'N,J!HU\!`FY>`FIE!
/+&!O'&J!-EJ!GHHIJ
! ! LITERATURVERZEICHNIS
GDH!
/0+$5)J!cEJ!CDD]J!L$02+5(!15(',+,!',!2'$)+2,!80$!'(25>M'$',5256!67+9027+$'MNz!<506759E!<50>M7N,E!K62'J!HUec\!GZZ>G`FE!
/0+$5)J!cEJ!c7'1$';'$25J!/EJ!W'MM+,J!<EJ!'(*!B'227+4,J!WEJ!CDDDJ![7+!6+&&!6N6&+!5(!M$020"0'(!M'$',52+,z!L$0)E!c+&&!cN6&+!P+,J!S\!G`Y>GFYE!
/0+$5)J!cEJ!/0+$5)J!cEJ!U0$$061,J!LEJ!c0N&+J!=EJ!c'$&20(J!=EJ!O#&2'(J!KEJ!K$(02J!/EJ!'(*!c'$2+$J!PEJ!GHHZJ!L86$1>GJ!'!*+@+&0M9+(2'&&N!$+)#&'2+*!6*6C>$+&'2+*!M$02+5(!15(',+!08!L&',90>*5#9!8'&65M'$#9z!B0&!<5067+9!L'$',520&J!cO\!G`I>GI]E!
/0+$5)J!cEJ!%(*56022J!=EJ!'(*!c7'1$';'$25J!/EJ!CDDCJ!cN6&5(>*+M+(*+(2!15(',+!7090&0)#+,!08!L&',90*5#9!8'&65M'$#9z!X(2!=!L'$',520&J!PJ\!GZIZ>GZFZE!
/0$5)0J!BEJ!B'(5+""0J!REJ!'(*!c0&0$(5J!KEJ!GHH`J!K(2!,N,2+9z!0M2595"'250(!;N!'!60&0(N!08!600M+$'25()!')+(2,z!X%%%![$'(,!ON,2!B'(E!cN;+$(E!<!cN;+$(EJ!JU\!CH>]GE!
/$#1+$J!<E!=EJ!CDDCJ!O[XZIG!e-&++@+6f!',!'!M'$'*5)9!80$!6'(6+$!27+$'MNz![$+(*,!B0&!B+*J!R\!OG]>OGFE!
/#;5(5('J!-E!-EJ!c7#M$N('J!dE!dEJ!L&'20(0@J!BE!dEJ!<0$5,10J!LE!dEJ!d,2$0@,1'J!-E!REJ![0&>9'670@J!KE!dEJ!'(*!O725&J!KE!KEJ!CDDIJ!X(!,5&560!*+,5)(!08!M$02+5(!15(',+!5(75;520$,z!,#6>6+,,+,!'(*!8'5&#$+,z!K(256'(6+$!K)+(2,!B+*!c7+9J!c\!GIG>GFFE!
/#*'J!PE!dEJ!U'$2J!LE!%EJ!'(*!O20$1J!/E!-EJ!CDDGJ!L'22+$(!c&',,5856'250(Ez!=07(!_5&+N!'(*!O0(,EJ!JE!
%**NJ!OE!PEJ!CDD]J!_7'2!5,!'!75**+(!B'$10@!90*+&éz!3'2!<502+67(0&EJ!JJ\!GYGZ>GYG`E!
%)&+(J!PE!BE!'(*!P+5,5(+J![EJ!CDDHJ![7+!6#$$+(2!,2'2#,!08!*$#)!*5,60@+$N!')'5(,2!27+!7#9'(!15(09+z!K,,'N!/$#)!/+@E![+67(0&J!c\!CC>]YE!
+&!U'4',7J!OE!KEJ!<'*'4+NJ!+EJ!'(*!W'MM+J![EJ!GHHHJ!<+("595*'"0&+!60(*+(,+*!$5()!,N,2+9,E!}XXE!ON(27+,5,!'(*!'(256'(6+$!+@'&#'250(!08!6+$2'5(!MN$5*0xGJC>'y;+("595*'"0&+!*+$5@'>25@+,z!L7'$9'"5+J!TS\!Y]G>Y]`E!
%&*$5*)+J!BE!/EJ!B#$$'NJ!cE!_EJ!K#20(J![E!PEJ!L'0&5(5J!-E!REJ!'(*!B++J!PE!LEJ!GHHIJ!%9M5$56'&!,60$5()!8#(6250(,z!XE![7+!*+@+&0M9+(2!08!'!8',2!+9M5$56'&!,60$5()!8#(6250(!20!+,259'2+!27+!;5(*5()!'885(52N!08!&5)'(*,!5(!$+6+M20$!609M&+\+,z!=!c09M#2E!K5*+*!B0&!/+,J!HH\!]CZ>]]ZE!
%&&+*)+J!OE!=EJ!GHH`J!c+&&!6N6&+!67+61M05(2,z!M$+@+(25()!'(!5*+(252N!6$5,5,z!O65+(6+J!JcS\!G``]>G`ICE!
%B%KJ!CDDHJ!722Mzoo444E+9+'E!+#$0M'E+#oE!
%(*56022J!=E!KEJ!30;&+J!BE!%EJ!'(*![#61+$J!=E!KEJ!GHHHJ!cN6&5(>*+M+(*+(2!15(',+,z!5(75;5250(!'(*!,#;,2$'2+!$+60)(5250(z!c#$$!dM5(!O2$#62E!<50&J!e\!IYF>I]]E!
%()+&,J!WEJ!CDDZJ
LITERATURVERZEICHNIS
GGD!
%()&5,7J!3E!=EJ!CDDIJ!c'&6#&'250(!08!;5(*5()!'885(525+,!08!UXR>G!P[!'(*!;+2'>,+6$+2',+!5(75;5>20$,!#,5()!27+!&5(+'$!5(2+$'6250(!+(+$)N!9+270*!4527!+\M&5652!'(*!60(25(##9!,0&@'250(!'MM$0'67+,z!=!B0&!B0*+&J!HP\!GDFG>GDHIE!
:';5'(J!BE!KEJ!<5)),J!_E!UEJ!XXXJ![$+5;+$J!/E!WEJ!K22+$5*)+J!cE!%EJ!K"5950'$'J!BE!/EJ!<+(+*+225J!BE!-EJ!c'$2+$J![E!KEJ!c56+$5J!LEJ!%*++(J!LE![EJ!:&0N*J!BEJ!:0$*J!=E!BEJ!-'&@5(J!BEJ!-+$&'67J!=E!VEJ!-$02"8+&*J!PE!BEJ!U+$$)'$*J!OEJ!X(,10J!/E!%EJ!X(,10J!BE!KEJ!V'5J!KE!-EJ!V+&5',J!=E!BEJ!B+72'J!OE!KEJ!B5&'(0@J!^E!REJ!R+&',60J!KE!BEJ!_0*561'J!VE!BEJ!L'2+&J!UE!WEJ!^'$$5(1'$J!LE!LEJ!'(*!V0617'$2J!/E!=EJ!CDDZJ!K!,9'&&!90&+6#&+>15(',+!5(2+$'6250(!9'M!80$!6&5(56'&!15(',+!5(75;520$,z!3'2!<502+67(0&EJ!JP\!YCH>YY`E!
:+$;NJ!XEJ!<&'"b#+"J!BEJ!L'&9+$J!KEJ!%$52p'J!PEJ!'(*!3+;$+*'J!KE!PEJ!GHHHJ!K!(0@+&!MY]e6*6Cf>;5(*5()!'(*!'625@'25()!M$02+5(!27'2!5,!(+6+,,'$N!'(*!,#88565+(2!20!2$5))+$!-eCfoB!M$0)$+,>,50(!5(!}+(0M#,!006N2+,z!-+(+,!/+@EJ!HP\!CGII>CGFHE!
:567+$'J!BE!%E!'(*!P00,J!/E!OEJ!GHHIJ!K!M&',25*!0$)'(+&&+!',!'!*$#)!2'$)+2!5(!'M5609M&+\'(!M'$',52+,z!3'2#$+J!PeO\!]DI>]DHE!
:5,67+$J!%EJ!GFH]J!%5(8&#,,!*+$!c0(85)#$'250(!'#8!*5+!_5$1#()!*+$!%("N9+Ez!<+$E!/2,67E!c7+9E!-+,EJ!Jc\!CHHYE!
:5,7+$J!PE!LEJ!CDDZJ!O+6$+2,!08!'!*0#;&+!')+(2z!c/WI!5(!6+&&>6N6&+!60(2$0&!'(*!2$'(,6$5M250(z!=!c+&&!O65J!HHR\!ZGIG>ZGFDE!
:0&0MM+J!3E!'(*!U#;;'$*J!PEJ!CDD`J![04'$*,!M$+*5625@+!&5)'(*!*+,5)(!4527!8$++>+(+$)N!;',+*!609M#2'250('&!9+270*,éz!c#$$!B+*!c7+9J!HP\!YZFY>Y`DFE!
:0$,2+$J!BE!=EJ!CDDCJ!B0&+6#&'$!90*+&&5()!5(!,2$#62#$'&!;50&0)Nz!B56$0(EJ!PP\!Y`Z>YF]E!
:027J!<E!=E!'(*!B6:'**+(J!-E!XEJ!CDDYJ![7+!'M560M&',2z!'!M&',25*!5(!L&',90*5#9!8'&65M'$#9!'(*!027+$!KM5609M&+\'(!M'$',52+,z!X(2!P+@!cN20&J!JJS\!ZI>GGDE!
:$'9M20(J!=E!%EJ!CDDHJ!V'M'25(5;z!'!$+@5+4!08!52,!#,+!5(!27+!2$+'29+(2!08!U%PC>0@+$+\M$+,,5()J!2$',2#"#9';>$+8$'620$NJ!'*@'(6+*!0$!9+2',2'256!;$+',2!6'(6+$z!/$#),J!Ue\!CGCZ>CG]FE!
-'0J!UE!}E!'(*!L+()J!VE!BEJ!GHHHJ!L'$'9+2+$5"'250(!08!27+!2+9M+$'2#$+!*+M+(*+(6+!08!27+!/+;N+>_'&&+$!8'620$,z!K62'!c$N,2'&&0)$E!KJ!TT\!HC`>HYCE!
-'$*(+$J!BE!=EJ!U'&&J!3EJ!:#()J!%EJ!_752+J!dEJ!<+$$59'(J!BEJ!UN9'(J!PE!_EJ!c'$&20(J!=E!BEJ!L'5(J!KEJ!3+&,0(J!WE!%EJ!<049'(J!OEJ!L'#&,+(J!XE![EJ!='9+,J!WEJ!%5,+(J!=E!KEJ!P#27+$80$*J!WEJ!O'&";+$)J!OE!VEJ!c$'5)J!KEJ!WN+,J!OEJ!c7'(J!BE!OEJ!3+(+J!REJ!O7'&&09J!OE!=EJ!O#7J!<EJ!L+2+$,0(J!=EJ!K()5#0&5J!OEJ!L+$2+'J!BEJ!K&&+(J!=EJ!O+&+()#2J!=EJ!U'82J!/EJ!B'27+$J!BE!_EJ!R'5*N'J!KE!<EJ!B'$25(J!/E!BEJ!:'5$&'9;J!KE!UEJ!:$'#(70&"J!BE!=EJ!P00,J!/E!OEJ!P'&M7J!OE!KEJ!B6:'**+(J!-E!XEJ!c#995(),J!VE!BEJ!O#;$'9'(5'(J!-E!BEJ!B#()'&&J!cEJ!R+(2+$J!=E!cEJ!c'$#665J!/E!=EJ!U0889'(J!OE!VEJ!3+4;0&*J!cEJ!/'@5,J!PE!_EJ!:$',+$J!cE!BEJ!'(*!<'$$+&&J!<EJ!CDDCJ!-+(09+!,+b#+(6+!08!27+!7#9'(!9'&'$5'!M'$',52+!L&',90*5#9!8'&65M'$#9z!3'>2#$+J!SHe\!]HF>ZGGE!
-+N+$J!=E!KEJ!L$5))+J!OE
! ! LITERATURVERZEICHNIS
GGG!
-5;,0(J!KE!%EJ!K$$5,J!cE!%EJ!<+(2&+NJ!=EJ!<0N&+J!:E![EJ!c#$25(J!3E!=EJ!/'@5+,J![E!-EJ!%(*56022J!=E!KEJ!-0&*5()J!<E![EJ!-$'(2J!OEJ!-$5885(J!PE!=EJ!=+4,;#$NJ!LEJ!=07(,0(J!VE!3EJ!B+,)#567+J!REJ!3+4+&&J!/E!PEJ!30;&+J!BE!%EJ![#61+$J!=E!KEJ!'(*!_75285+&*J!UE!=EJ!CDDCJ!L$0;5()!27+!K[L!$5;0,+>;5(*5()!*09'5(!08!6N6&5(>*+M+(*+(2!15(',+,!G!'(*!C!4527!de`f>,#;,252#2+*!)#'(5(+!*+$5@'25@+,z!=!B+*!c7+9J!ST\!YYFG>YYHYE!
-0$$+J!BE!%EJ!B07'99+*J!BEJ!%&&400*J!WEJ!U,#J!3EJ!L'b#+22+J!PEJ!P'0J!LE!3EJ!'(*!O'4N+$,J!cE!VEJ!CDDGJ!c&5(56'&!$+,5,2'(6+!20!O[X>ZIG!6'(6+$!27+$'MN!6'#,+*!;N!<cP>K<V!)+(+!9#2'250(!0$!'9M&5856'250(z!O65+(6+J!JeP\!FI`>FFDE!
-$'(2J!WE!BEJ!/#(50(J!BE!UEJ!j'$*&+NJ!REJ!O1'&2,0#(5,J!KE!VEJ!B'$10J!/EJ!%5,+(;$'(*J!-EJ!c$082J!OE!VEJ!B+5p+$J!VEJ!'(*!B022$'9J!=E!cEJ!CDD]J!X(75;520$,!08!V+5,79'(5'!9+\56'('!cPWY!6N6&5(>*+M+(*+(2!15(',+z!67+956'&!&5;$'$N!,6$++(!'(*!'(25&+5,79'(5'&!'625@52Nz!K(25956$0;!K)+(2,!c7+9027+$J!SR\!YDYY>YD]CE!
-$'NJ!3EJ!/+25@'#*J!VEJ!/0+$5)J!cEJ!'(*!B+5p+$J!VEJ!GHHHJ!K[L>,52+!*5$+62+*!5(75;520$,!08!6N>6&5(>*+M+(*+(2!15(',+,z!c#$$!B+*!c7+9J!U\!FZH>FIZE!
U'&)$+(J![E!KEJ!GHH`J!B+$61!B0&+6#&'$!:0$6+!:5+&*E!X>REz!=E!c09M#2E!c7+9EJ!Hc\!]HD>`]GE!
U'&&J![E!KEJ!CDDHJ!<50%*52z!'!#,+$>8$5+(*&N!;50&0)56'&!,+b#+(6+!'&5)(9+(2!+*520$!'(*!'('&N,5,!M$0)$'9!80$!_5(*04,!HZoHFo3[Ez!3#6&E!K65*,E!ON9ME!O+$EJ!HZ>HFE!
U'99'$20(J![E!cEJ!B022$'9J!=E!cEJ!'(*!/0+$5)J!cEJ!CDDYJ![7+!6+&&!6N6&+!08!M'$',5256!M$020"0'z!M02+(25'&!80$!67+9027+$'M+#256!+\M&052'250(z!L$0)E!c+&&!cN6&+!P+,J!T\!HG>GDGE!
U'(1,J!OE!WE!'(*!U#(2+$J![EJ!GHHZJ!L$02+5(!15(',+,!`E![7+!+#1'$N0256!M$02+5(!15(',+!,#M+$>8'95&Nz!15(',+!e6'2'&N256f!*09'5(!,2$#62#$+!'(*!6&',,5856'250(z!:KO%<!=J!e\!ZI`>ZH`E!
U'$9,+J!VEJ!@'(!^N&J!PEJ!-$'NJ!3EJ!O67#&2"J!LEJ!V+6&+$6J!OEJ!B+5p+$J!VEJ!/0+$5)J!cEJ!'(*!U'@&51J!XEJ!CDDGJ!O2$#62#$+>'625@52N!$+&'250(,75M,!'(*!5(75;520$N!+88+62,!08!@'$50#,!M#$5(+!*+$5@'>25@+,!0(!27+!5(!@52$0!)$0427!08!L&',90*5#9!8'&65M'$#9z!<5067+9!L7'$9'60&J!UJ\!Y]G>Y]FE!
U',,'(J!LEJ!:+$)#,,0(J!/EJ!-$'(2J!WE!BEJ!'(*!B022$'9J!=E!cEJ!CDDGJ![7+!cPWY!M$02+5(!15(',+!5,!+,,+(25'&!80$!6+&&!6N6&+!M$0)$+,,50(!08!V+5,79'(5'!9+\56'('z!B0&!<5067+9!L'$',520&J!HHP\!GFH>GHFE!
U'#,675&*J!KEJ!CDDIJ!c&5(56'&!,2#*5+,!4527!,0$'8+(5;!e3+\'@'$f!5(!9+2',2'256!9+&'(09'z!=!/2,67E!/+$9'20&!-+,J!T\!Y]ZE!
U+(51088J!OE!'(*!U+(51088J!=E!-EJ!GHHYJ!L+$80$9'(6+!+@'&#'250(!08!'95(0!'65*!,#;,252#250(!9'2$56+,z!L$02+5(,J!Hc\!]H>`GE!
U067+))+$J!UEJ
LITERATURVERZEICHNIS
GGC!
U#'()J!3EJ!O70567+2J!<E!WEJ!'(*!X$45(J!=E!=EJ!CDD`J!<+(679'$15()!,+2,!80$!90&+6#&'$!*0615()z!=!B+*!c7+9J!Se\!`IFH>`FDGE!
U#&0J!3EJ!<'5$067J!KEJ!<#&&5'$*J!REJ!c+$#225J!VEJ!*+!c',2$0J!%EJ!V'()+(*5p1>-+(+@'#\J!LE!OEJ!L')(5J!BEJ!'(*!O5)$5,2J!cE!=EJ!CDD`J![7+!LPdOX[%!*'2';',+z!3#6&+56!K65*,!P+,J!PS\!/CCI>/CYDE!
XU-OcJ!CDD]J!:5(5,75()!27+!+#67$09'256!,+b#+(6+!08!27+!7#9'(!)+(09+z!3'2#$+J!SPH\!HYG>H]ZE!
X7&+(8+&*2J!_E!/EJ!R05)2J!=E!UEJ!<5+(8'52J!<EJ!d+&&5+(J!:EJ!'(*!3561&'#,J!BE!cEJ!CDDCJ!%(7'(6+*!cKc[RO!;$04,+$!08!27+!dM+(!3cX!/'2';',+z!=!c7+9!X(8!c09M#2E!O65J!SJ\!]`>ZIE!
Xb;'&J!WEJ!K&0(,0J!KE!cEJ!%&!K11'*J!%EJ!-0()J!cE!}EJ!U'b#+J!3EJ!W7'200(J!OEJ!L+5J!=E!=EJ!['(5>9#1'5J!UEJ![,#p50J!XEJ!_'()J!=E!^EJ!'(*!-$#(*1+>Xb;'J!XEJ!CDDYJ!K&"7+59+$!(+#$085;$5&&'$N!*+)+(+$'250(z!27+$'M+#256!2'$)+2,!'(*!75)7>27$0#)7M#2!',,'N,z!=!B0&!3+#$0,65J!JO\!]CZ>]CHE!
='66'$*J!LEJ!GHGCJ![7+!*5,2$5;#250(!08!8&0$'!5(!27+!'&M5(+!"0(+z!3+4!L7N20&0)5,2J!HH\!YI>ZDE!
='5(J!KE!3EJ!GHH`J!O60$5()!(0(60@'&+(2!M$02+5(>&5)'(*!5(2+$'6250(,z!'!60(25(#0#,!*588+$+(25';&+!8#(6250(!2#(+*!20!609M#2+!;5(*5()!'885(525+,z!=!c09M#2E!K5*+*!B0&!/+,J!HO\!]CI>]]DE!
='#2+&'2J!PEJ!<$#9;NJ![EJ!O67'8+$J!BEJ!<$5+9J!UEJ!%5,+(;$'(*J!-EJ!O674'7(J!OEJ!W$#)+$J!BEJ!V#615()J!?EJ!L$5+(J!dEJ!'(*!O5+9+5,2+$J!-EJ!CDDZJ!:$09!27+!5(,0&#;&+!*N+!5(*5$#;5(!20>4'$*,!75)7&N!'625@+J!,0&#;&+!c/WC>5(75;520$,z!c7+9;5067+9J!U\!ZYG>Z]DE!
=+88$+NJ!LE!/EJ!P#,,0J!KE!KEJ!L0&N'1J!WEJ!-5;;,J!%EJ!U#$452"J!=EJ!B',,')#+J!=EJ!'(*!L'@&+2567J!3E!LEJ!GHHZJ!B+67'(5,9!08!c/W!'625@'250(!$+@+'&+*!;N!27+!,2$#62#$+!08!'!6N6&5(K>c/WC!609M&+\z!3'2#$+J!PcU\!YGY>YCDE!
=5'()J!^EJ!CDDFJ!c09M#2'250('&!'('&N,5,!08!27+!5(2+$'6250(!;+24++(!&5)'(*>$+6+M20$!M'5$,z!c#$$!L7'$9!/+,J!HS\!ZFF>ZHCE!
=07'(,,0(J!BE!'(*!L+$,,0(J!=E!VEJ!CDDFJ!c'(6+$!27+$'MNz!2'$)+25()!6+&&!6N6&+!$+)#&'20$,z!K(25>6'(6+$!K)+(2,!B+*!c7+9J!R\!ICY>IYGE!
=07(,0(J!/E!-E!'(*!_'&1+$J!cE!VEJ!GHHHJ!cN6&5(,!'(*!6+&&!6N6&+!67+61M05(2,z!K((#E!P+@!L7'$9'60&![0\560&J!Pe\!CHZ>YGCE!
=07(,0(J!VE!3EJ!CDDHJ!L$02+5(!15(',+!5(75;520$,z!60(2$5;#250(,!8$09!,2$#62#$+!20!6&5(56'&!609>M0#(*,z!iE!P+@!<50M7N,EJ!SJ\!G>]DE!
=09''J!UEJ!_5+,(+$J!=EJ!O'(*+$;$'(*J!OEJ!K&25(656+1J!<EJ!_+5*+9+N+$J!cEJ!U5(2"J!BEJ![#$>;'670@'J!XEJ!%;+$&J!BEJ!^+5*&+$J!=EJ!V5672+(27'&+$J!UE!WEJ!O0&*'25J!/EJ!'(*!<+61J!%EJ!GHHHJ!X(75;520$,!08!27+!(0(9+@'&0('2+!M'274'N!08!5,0M$+(05*!;50,N(27+,5,!',!'(259'&'$5'&!*$#),z!O65+(6+J!JRT\!GZIY>GZI`E!
=0(+,J!-EJ!_5&&+22J!LEJ!'(*!-&+(J!PE!cEJ!GHHZJ!B0&+6#&'$!$+60)(5250(!08!$+6+M20$!,52+,!#,5()!'!)+(+256!'&)0$5279!4527!'!*+,6$5M250(!08!*+,0&@'250(z!=!B0&!<50&J!JST\!]Y>ZYE!
! ! LITERATURVERZEICHNIS
GGY!
=0(+,J!-EJ!_5&&+22J!LEJ!-&+(J!PE!cEJ!V+'67J!KE!PEJ!'(*!['N&0$J!PEJ!GHHIJ!/+@+&0M9+(2!'(*!@'&5>*'250(!08!'!)+(+256!'&)0$5279!80$!8&+\5;&+!*0615()z!=!B0&!<50&J!JUc\!ICI>I]FE!
W'&*5,J!LEJ!GHHHJ![7+!6*1>'625@'25()!15(',+!ecKWfz!8$09!N+',2!20!9'99'&,z!c+&&!B0&!V58+!O65J!TT\!CF]>CH`E!
W'(2+$J!LEJ!CDDZJ!%5(+!%@'&#'250(!*+$!-+('#5)1+52!@+$,675+*+(+$!R+$8'7$+(!*+$!-+(@0$7+$,'>)+!5(!)$0+(!/3K>O+b#+("+(Ez!722Mzoo27+0$5+E5(80$9'251E#(5>#&9E!*+oV+7$+oOOZo-RoK#,'$;+52#()áW'(2+$E!M*8E!
W+&&+(;+$)+$J!%EJ!P0*$5)0J!=EJ!B#&&+$J!LEJ!'(*!P0)('(J!/EJ!CDD]J!c09M'$'25@+!+@'&#'250(!08!+5)72!*0615()!200&,!80$!*0615()!'(*!@5$2#'&!,6$++(5()!'66#$'6Nz!L$02+5(,J!Tc\!CCZ>C]CE!
W75$+J!?E!PEJ!<'(1,20(J!/EJ!<'$;0,'J!=EJ!<$522+&&5J!/E!PEJ!c'$5()'&J!jEJ!c'$&,0(J!PEJ!/#9',J!=EJ!-'(+J![EJ!U+'&*J!OE!VEJ!U5;(+$J!<EJ!=07(,0(J!=E!OEJ!W'2"J!BE!%EJ!W+((#$+J!3EJ!W5()+$N>_00*J!=EJ!V++J!_EJ!V5#J!}E!-EJ!V045()+$J![E!<EJ!B6K&+\'(*+$J!XEJ!B0('7'(J!BE!WEJ!3'2+$0J!PEJ!P+(561J!=EJ!P5+*&J!<EJ!P0()J!UEJ!O5;&+NJ!PE!3EJ!O9527J!PE!KEJ!'(*!_0&'(5(J!/EJ!CDD]J!d9+)'>6'$;0\NMN$5*N&!,#;,252#2+*!#$+',!',!P'8!15(',+!5(75;520$,z!OKP!08!27+!'9>5*+!,#;,252#+(2z!<500$)E!B+*!c7+9!V+22J!HS\!IFY>IF`E!
W59J!UE!jE!'(*!c70J!jEJ!GHHIJ!O2$#62#$'&!,595&'$52N!;+24++(!27+!M061+2!$+)50(!08!$+25(0;&',>209'!2#90#$!,#MM$+,,0$!'(*!27+!6N6&5(>;0\z!3'2!O2$#62E!<50&J!S\!YHD>YHZE!
W5(('5$*J!=EJ!V0)'(J!BEJ!['52J!KEJ!'(*!V'(),&+NJ!-EJ!CDDGJ!['cPWY!+(60*+,!'!(0@+&![7+5&+$5'!'((#&'2'!M$02+5(!15(',+!4527!90258,!67'$'62+$5,256!08!27+!8'95&N!08!+#1'$N0256!6N6&5(!*+>M+(*+(2!15(',+,z!'!609M'$'25@+!'('&N,5,!08!52,!+\M$+,,50(!4527!['cPWC!*#$5()!27+!M'$'>,52+!&58+!6N6&+z!-+(+J!Jce\!GCI>GYZE!
W5(('5$*J!=E!UEJ!<#9,2+'*J!=E!BEJ!B'((J!/E!=EJ!PN'(J!PEJ!O75$&+NJ!BE!_EJ!O75+&,J!<E!PEJ!'(*![09&+NJ!:E!BEJ!CDD]J!%2cPWCJ!'!6N6&5(>*+M+(*+(2!15(',+!)+(+!+\M$+,,+*!*#$5()!27+!,+\#'&!'(*!',+\#'&!M7',+,!08!27+!%59+$5'!2+(+&&'!&58+!6N6&+z!X(2E!=!L'$',520&EJ!PS\!`FY>`HCE!
W52')'4'J!BEJ!d1';+J![EJ!d)5(0J!UEJ!B'2,#9020J!UEJ!O#"#15>['1'7',75J!XEJ!W01#;0J![EJ!U5)'>,75J!UEJ!O'5207J!OEJ!['N'J!jEJ!j',#*'J!UEJ!'(*!EJ!GHHYJ!<#2N$0&'620(+!XJ!'!,+&+625@+!5(75;5>20$!08!6*1C!'(*!6*6C!15(',+z!d(60)+(+J!R\!C]CZ>C]YCE!
W5267+(J!/E!<EJ!/+60$(+"J!UEJ!:#$$J!=E!PEJ!'(*!<'p0$'27J!=EJ!CDD]J!/0615()!'(*!,60$5()!5(!@5$>2#'&!,6$++(5()!80$!*$#)!*5,60@+$Nz!9+270*,!'(*!'MM&56'250(,z!3'2!P+@!/$#)!/5,60@J!P\!HYZ>H]HE!
W&+;+J!-EJ!CDD`J!R5$2#'&!&5)'(*!,6$++(5()z!,2$'2+)5+,J!M+$,M+625@+,!'(*!&5952'250(,z!/$#)!/5,>60@![0*'NJ!HH\!ZFD>ZH]E!
W&+;+J!-EJ!CDDHJ!_5$1,2088*+,5)(z!%(24#$8!#(*!_5$1#()!@0(!K$"(+5,2088+(z!OM+12$#9!K1'*E!R+$&')J!U+5*+&;+$)J!<+$&5(J!d\80$*EJ!J\!]CI>]ZFE!
W&5(1+$2J!BE!iE!'(*!U+#,,&+$J!REJ!CDD`J![7+!#,+!08!'(256'(6+$!*$#),!5(!'(25M'$',5256!67+9027+>$'MNz!B5(5!P+@!B+*!c7+9J!U\!GYG>G]YE!
W(5)720(J!/E!PEJ!}#0()J!3E!UEJ
LITERATURVERZEICHNIS
GG]!
'95(0!'65*!$+,5*#+!M+M25*+!5(75;520$!'(*!B)K[L!*588$'62!20!YED!K!$+,0ú(z!=!B0&!<50&J!JJO\!CGI>CCDE!
W(##25&'J!OEJ!<p0$1b@5,2J!KE!BEJ!K#250J!WEJ!['$11'(+(J!BEJ!_0&8J!BEJ!B0((5J!dEJ!O"N9'(,1'J!=EJ!V'$$'9+(*NJ!BE!VEJ!['MM+$J!=EJ!L+$+J!UEJ!+&!P58'5J!_EJ!U+99+$J!OEJ!_',+(5#,J!RE!BEJ!R5*)$+(J!REJ!'(*!^7#J!jEJ!GHHFJ!/3K!60MN!(#9;+$!'9M&5856'250(,!5(!7#9'(!(+0M&',9,z!$+@5+4!08!609M'$'25@+!)+(0956!7N;$5*5"'250(!,2#*5+,z!K9!=!L'270&J!HTJ\!GGDI>GGCYE!
W0;'N',75J!UEJ!O2+4'$2J!%EJ!L00(J!PEJ!K*'96"+4,15J!=E!LEJ!-'((0(J!=EJ!'(*!U#(2J![EJ!GHHCJ!X*+(25856'250(!08!27+!*09'5(,!5(!6N6&5(!K!$+b#5$+*!80$!;5(*5()!20J!'(*!'625@'250(!08J!MY]6*6C!'(*!MYC6*1C!M$02+5(!15(',+!,#;#(52,z!B0&!<50&!c+&&J!P\!GCIH>GCH]E!
W0&;J!LEJ!U#'()J!/EJ!/+NJ!:EJ!'(*!c'8&5,67J!KEJ!CDDFJ!/5,60@+$N!08!15(',+!5(75;520$,!;N!75)7>27$0#)7M#2!*0615()!'(*!,60$5()!;',+*!0(!'!2$'(,8+$';&+!&5(+'$!5(2+$'6250(!+(+$)N!90*+&z!=!B+*!c7+9J!TH\!GGIH>GGFFE!
W0$;J!dEJ!O2Q2"&+J![EJ!'(*!%\(+$J![E!%EJ!CDD`J!O4'$9!X(2+&&5)+(6+E!LVK3[Oz!KMM&56'250(!08!K(2!c0&0(N!dM2595"'250(!20!O2$#62#$+><',+*!/$#)!/+,5)(z!OM$5()+$!R+$&')J!U+5*+&>;+$)J!<+$&5(J!3+4!j0$1J!H\!GGZ>GY]E!
W0$;J!dEJ!O2#2"&+J![EJ!'(*!%\(+$J![E!%EJ!CDDHJ!%9M5$56'&!,60$5()!8#(6250(,!80$!'*@'(6+*!M$0>2+5(>&5)'(*!*0615()!4527!LVK3[Oz!=!c7+9!X(8!B0*+&J!Se\!F]>H`E!
W0$8J!XEJ!CDDYJ!O+$5'&!<VKO[!,+'$675()z!<505(80$9'256,J!He\!G]HC>G]H`E!
W0$0,J!%E!'(*!/0$(+$>c50,,+1J!cEJ!CDDIJ![7+!$0&+!08!)&N60)+(!,N(27',+!15(',+>Y;+2'!5(!,675"0M7$+(5'z!/$#)!3+4,!L+$,M+62EJ!JO\!]YI>]]ZE!
W0,15J!VE!<E!'(*!-0&*5()J!-E!<EJ!CDDGJ![7+!6&0,+,2!<VKO[!752!5,!082+(!(02!27+!(+'$+,2!(+5)7;0$z!=!B0&!%@0&J!TJ\!Z]D>Z]CE!
W$+;,J!%E!-E!'(*!<+'@0J!=E!KEJ!GHIHJ!L70,M70$N&'250(>*+M70,M70$N&'250(!08!+("N9+,z!K((#E!P+@!<5067+9J!SR\!HCY>HZHE!
W$0+9+$J!PE![EJ!CDDIJ!O2$#62#$+>;',+*!*$#)!*+,5)(z!*0615()!'(*!,60$5()z!c#$$!L$02+5(!L+M2E!O65J!R\!YGC>YCFE!
W$N,72'80@N67J!KEJ!L$&56J!KEJ!/9N2$5@J!^EJ!/'(5J!LEJ!B5&0,2'(J!BEJ!%N$567J!REJ!U#;;'$*J![EJ!'(*!:5*+&5,J!WEJ!CDDIJ!3+4!200&,!'(*!+\M'(*+*!*'2'!'('&N,5,!6'M';5&525+,!'2!27+!L$02+5(!O2$#62#$+!L$+*56250(!c+(2+$z!L$02+5(,J!Ue\!GH>C`E!
W#;5(N5J!UE!'(*!BQ&&+$J!-EJ!CDD]J!c7+90)+(0956,!5(!/$#)!/5,60@+$NE!K!B+*565('&!c7+>95,2$N!L+$,M+625@+z!_5&+N>RcU!R+$&')J!_+5(7+59J!H\!GHG>CGHE!
W#99+$8+&*J!3EJ!CDDZJ!W$'(17+52+(!*+$!U+5925+$+.!W'M52+&!GFz!UQ7(+$@g)+&z!O67&Q2+$,67+!R+$&'),)+,+&&,67'82!9;U!k!c0E!W-J!U\!`GY>`ZGE!
W@''&J!cE!KEJ!P'*1+J!=E!PEJ!-#+$5(5J!BE!3EJ!'(*!_752+J!BE!_EJ!CDDCJ!X,0&'250(!08!'![0\0M&',9'!)0(*55!6N6&5(!;N!N+',2!240>7N;$5*!5(2+$'625@+!,6$++(z!B0&!<5067+9!L'$',5>20&J!HJO\!GFI>GH]E!
! ! LITERATURVERZEICHNIS
GGZ!
V'()+J!KEJ!B6V'(+J!VE!BEJ!B5&&,J!PE!%EJ!/+@5(+J!OE!%EJ!'(*!c0$;+22J!KE!UEJ!CDDHJ!%\M'(*5()!27+!/+85(5250(!08!27+!c&',,56'&!<5M'$252+!3#6&+'$!V06'&5"'250(!O5)('&z![$'8856EE!
V+!P067J!WEJ!O+,25+$J!cEJ!/0$5(J!/EJ!_'2+$,J!3EJ!W'MM+,J!<EJ!c7'1$';'$25J!/EJ!B+5p+$J!VEJ!'(*!/0+$5)J!cEJ!CDDDJ!K625@'250(!08!'!L&',90*5#9!8'&65M'$#9!6*6C>$+&'2+*!15(',+!;N!7+2>+$0&0)0#,!MCZ!'(*!6N6&5(!UE!:#(6250('&!67'$'62+$5"'250(!08!'!LE!8'&65M'$#9!6N6&5(!7090>&0)#+z!=!<50&!c7+9J!JcT\!FHZC>FHZFE!
V+'67J!KE!PEJ!O70567+2J!<E!WEJ!'(*!L+5,7088J!cE!%EJ!CDD`J!L$+*56250(!08!M$02+5(>&5)'(*!5(2+$'6>250(,E!/0615()!'(*!,60$5()z!,#66+,,+,!'(*!)'M,z!=!B+*!c7+9J!Se\!ZFZG>ZFZZE!
V++,J!%E!BE!'(*!U'$&04J!%EJ!GHHYJ!O+b#+(6+,!45275(!27+!60(,+$@+*!6N6&5(!;0\!08!7#9'(!6N>6&5(!K!'$+!,#88565+(2!80$!;5(*5()!20!'(*!'625@'250(!08!6*6C!15(',+z!B0&!c+&&!<50&J!HP\!GGH]>GCDGE!
V+45(J!<EJ!CDDFJ!-+(+,!X}z!=0(+,!'(*!<'$2&+22!L#;&5,7+$,J!X(6E!J!V0(*0(J!?WE!
V5J!=E!VEJ!P0;,0(J!WE!=EJ!c7+(J!=E!VEJ!['$)+22J!-E!KEJ!'(*!<'1+$J!/E!KEJ!GHH`J!L89$1J!'!BdGZ>$+&'2+*!M$02+5(!15(',+!8$09!L&',90*5#9!8'&65M'$#9E!-+(+!6&0(5()J!,+b#+(6+J!,2')+>,M+65856!+\M$+,,50(!'(*!67$090,09+!&06'&5"'250(z!%#$!=!<5067+9J!JSH\!FDZ>FGYE!
V5(*;+$)J!PE!KEJ!i#5((J!KE!BEJ!'(*!U#(2+$J![EJ!GHHCJ!/#'&>,M+6585652N!M$02+5(!15(',+,z!45&&!'(N!7N*$0\N&!*0éz![$+(*,!<5067+9!O65J!Hc\!GG]>GGHE!
V5#J!=E!=EJ!/+$9'2'15,J!KEJ!V#1'6,J!cEJ!W0("+&9'((J!:EJ!c7+(J!jEJ!W'99&022J!?EJ!/+M5(20J!_EJ!j'()J!UEJ!j5(J!}EJ!c7+(J!jEJ!O67#22J!KEJ!O5960\J!BE!%EJ!'(*!V#1J!WE!cEJ!CDDYJ!YJZJ`>[$5,#;,252#2+*!('M7270,2N$5&,!',!c/WC!5(75;520$,z!<500$)E!B+*!c7+9!V+22J!HP\!C]`Z>C]`FE!
V5#J![EJ!V5(J!jEJ!_+(J!}EJ!=0$5,,+(J!PE!3EJ!'(*!-5&,0(J!BE!WEJ!CDDIJ!<5(*5()/<z!'!4+;>'66+,,5;&+!*'2';',+!08!+\M+$59+(2'&&N!*+2+$95(+*!M$02+5(>&5)'(*!;5(*5()!'885(525+,z!3#>6&+56!K65*,!P+,J!PT\!/GHF>/CDGE!
V#65#,J!PE!'(*!V00,>:$'(1J!<EJ!CDDFJ!<50&0)5+!@0(!L'$',52+(Ez!OM$5()+$!R+$&')J!<+$&5(J!U+5>*+&;+$)J!3+4!j0$1J!GDI>CZ`E!
B'6*0('&*J!VEJ!CDDDJ!3+4!5(8&#+("'!*$#),!"'('95@5$!eP+&+("'f!'(*!0,+&2'95@5$!e['958&#fz!#(+\M+62+*!,+$50#,!$+'6250(,z!cBK=EJ!HUP\!FIH>ZE!
B'&5""5'J!VE!=E!'(*!U,#J!KEJ!CDDFJ![+9,5$0&59#,J!'(!9[dP!5(75;520$!80$!2$+'29+(2!08!M'25+(2,!4527!'*@'(6+*!$+('&!6+&&!6'$65(09'z!c&5(!=!d(60&E!3#$,EJ!HJ\!`YH>`]`E!
B'	$+,J!BEJ!CDDZJ!P+@5,525()!27+!.c*1>6+(2$56.!@5+4!08!27+!9'99'&5'(!6+&&!6N6&+z!c+&&!cN6&+J!S\!CD`>CGDE!
B'	$+,J!BE!'(*!<'$;'65*J!BEJ!CDDZJ!B'99'&5'(!6N6&5(>*+M+(*+(2!15(',+,z![$+(*,!<50>67+9!O65J!PO\!`YD>`]GE!
B'	$+,J!BE!'(*!<'$;'65*J!BEJ!CDDHJ!c+&&!6N6&+J!c/W,!'(*!6'(6+$z!'!67'()5()!M'$'*5)9z!3'2!P+@!c'(6+$J!e\!GZY>G``E!
LITERATURVERZEICHNIS
GG`!
B'((5()J!-EJ!_7N2+J!/E!<EJ!B'$25(+"J!PEJ!U#(2+$J![EJ!'(*!O#*'$,'('9J!OEJ!CDDCJ![7+!M$02+5(!15(',+!609M&+9+(2!08!27+!7#9'(!)+(09+z!O65+(6+J!JeR\!GHGC>GHY]E!
B6K&&5,2+$J!BE!BEJ!CDDZJ!K!*+6'*+!08!*5,60@+$5+,!5(!@+2+$5('$N!M$020"00&0)N!67'()+,!0#$!60(6+M2!08!.,#;6&5(56'&.!20\0M&',90,5,z!R+2!L'$',520&J!HPJ\!C]G>C]IE!
B6/0('&*J!RE!'(*!O75$&+NJ!BE!_EJ!CDDHJ!L',2!'(*!8#2#$+z!@'665('250(!')'5(,2!%59+$5'z!L'$',5>20&0)NJ!HPU\!G]II>G]FHE!
B6X((+,J!cEJ!CDDIJ!R5$2#'&!,6$++(5()!,2$'2+)5+,!5(!*$#)!*5,60@+$Nz!c#$$!dM5(!c7+9!<50&J!HH\!]H]>ZDCE!
B6X((+,J!cE!'(*!:5,67+$J!LE!BEJ!CDDZJ!O2$'2+)5+,!80$!27+!*+,5)(!08!M02+(2!'(*!,+&+625@+!15(',+!5(75;520$,z!c#$$!L7'$9!/+,J!HH\!GF]Z>GF`YE!
B+7&70$(J!UEJ!CDDGJ!%(6N6&0M+*56!$+8+$+(6+!08!M'$',520&0)Nz!M$5()+$!R+$&')J!<+$&5(J!U+5*+&>;+$)J!3+4!j0$1E!
B+7&70$(J!UEJ!CDDFJ!%(6N6&0M+*5'!08!M'$',520&0)Nz!OM$5()+$!R+$&')J!<+$&5(J!U+5*+&;+$)J!3+4!j0$1E!
B+7&70$(J!UEJ!/Q4+&J!/EJ!'(*!P'+27+$J!_EJ!GHHYJ!/5')(0,+!#(*![7+$'M5+!*+$!L'$',520,+(!@0(!U'#,>J!3#2">!#(*!U+5925+$+(z!-#,2'@!:5,67+$!R+$&')J!O2#22)'$2J!=+('J!3+4!j0$1J!YDY>Y`FE!
B+5+$J!?E![E!'(*!<&0;+&J!-EJ!GHHDJ!K!(#6&+'$!&06'&5"'250(!,5)('&!;5(*5()!M$02+5(!5(!27+!(#6&+0>&#,z!=!c+&&!<50&J!HHH\!CCYZ>CC]ZE!
B+5p+$J!VEJ!<0$)(+J!KEJ!B#&(+$J!dEJ!c70()J!=E!LEJ!<&04J!=E!=EJ!X(')'15J!3EJ!X(')'15J!BEJ!/+&>6$0,J!=E!-EJ!'(*!B0#&5(0#\J!=E!LEJ!GHHIJ!<5067+956'&!'(*!6+&&#&'$!+88+62,!08!$0,60@525(+J!'!M02+(2!'(*!,+&+625@+!5(75;520$!08!27+!6N6&5(>*+M+(*+(2!15(',+,!6*6CJ!6*1C!'(*!6*1Zz!%#$!=!<5067+9J!JSP\!ZCI>ZY`E!
B+&+(*+"J!PE!/EJ!CDDDJ!:#2#$+!M+$,M+625@+,!0(!@+2+$5('$N!7+90M'$',52+!$+,+'$67!5(!27+!2$0M56,!'2!27+!,2'$2!08!275,!6+(2#$Nz!K((!3!j!K6'*!O65J!eHU\!CZY>CZFE!
B+$61\J!KEJ!V+!P067J!WEJ!35@+"J!BE!LEJ!/0$5(J!/EJ!K&'(0J!LEJ!-#25+$$+"J!-E!=EJ!3+;$+*'J!KE!PEJ!-0&*$5()J!/EJ!_7522&+J!cEJ!L'22+$,0(J!OEJ!c7'1$';'$25J!/EJ!'(*!/0+$5)J!cEJ!CDDYJ!X*+(2585>6'250(!'(*!5(525'&!67'$'62+$5"'250(!08!27$++!(0@+&!6N6&5(>$+&'2+*!M$02+5(,!08!27+!7#9'(!9'&'$5'!M'$',52+!L&',90*5#9!8'&65M'$#9z!=!<50&!c7+9J!JcR\!YHFYH>YHFZDE!
B0('60J!%E!KE!'(*!R'&&'(0J!BE!VEJ!CDDYJ!cN6&5(>*+M+(*+(2!15(',+!5(75;520$,z!6'(6+$!15&&+$,!20!(+#$0('&!)#'$*5'(,z!c#$$!B+*!c7+9J!HO\!Y`I>YIHE!
B0(20N'J!=E!-E!'(*!V5+,+(8+&*J!dEJ!CDD]J
! ! LITERATURVERZEICHNIS
GGI!
B022$'9J!=E!cEJ!B6c$+'*NJ!<E!LEJ!<$04(J!WE!-EJ!'(*!-$'(2J!WE!BEJ!GHH`J!-+(+!*5,$#M250(,!5(*56'2+!'(!+,,+(25'&!8#(6250(!80$!27+!V99cPWG!6*6C>$+&'2+*!15(',+!08!V+5,79'(5'!9+\56'('z!B0&!B56$0;50&J!JJ\!ZIY>ZFYE!
B0#&2J!=EJ!:5*+&5,J!WEJ!^+9&'J!KEJ!'(*!U#;;'$*J![EJ!CDDGJ!c$5256'&!',,+,,9+(2!08!9+270*,!08!M$02+5(!,2$#62#$+!M$+*56250(!ecKOLfz!$0#(*!XRz!L$02+5(,J!T\!C>IE!
B0#(2J!OE!BEJ!GHFCJ!K!6'2'&0)#+!08!,M&56+!p#(6250(!,+b#+(6+,z!3#6&+56!K65*,!P+,J!HO\!]ZH>]ICE!
B#&*+$J!3E!=EJ!KM4+5&+$J!PEJ!K22400*J![E!WEJ!<'5$067J!KEJ!<'2+9'(J!KEJ!<5((,J!/EJ!<$'*&+NJ!LEJ!<0$1J!LEJ!<#67+$J!LEJ!c+$#225J!VEJ!c0M&+NJ!PEJ!c0#$6+&&+J!%EJ!/',J!?EJ!/#$;5(J!PEJ!:&+5,679'((J!_EJ!-0#)7J!=EJ!U'82J!/EJ!U'$2+J!3EJ!U#&0J!3EJ!W'7(J!/EJ!W'('M5(J!KEJ!W$+,2N'(5(0@'J!BEJ!V0(,*'&+J!/EJ!V0M+"J!PEJ!V+2#(56J!XEJ!B'*+$'J!BEJ!B',&+(J!=EJ!B6/04'&&J!=EJ!B5267+&&J!KEJ!3510&,1'N'J!KE!3EJ!d$67'$*J!OEJ!L')(5J!BEJ!L0(25()J!cE!LEJ!i#+@5&&0(J!%EJ!O+&+()#2J!=EJ!O5)$5,2J!cE!=EJ!O5&@+(205(+(J!REJ!O2#*70&9+J!/E!=EJ!R'#)7'(J!PEJ!'(*!_#J!cE!UEJ!CDDZJ!X(2+$L$0J!M$0)$+,,!'(*!,2'2#,!5(!CDDZz!3#6&+56!K65*,!P+,J!PP\!/CDG>/CDZE!
B#$M7NJ!OE!cE!'(*!<$+9'(J!=E!-EJ!CDDGJ!-'M,!5(!27+!675&*700*!9'&'$5'!;#$*+(!5(!K8$56'z!6+$+>;$'&!9'&'$5'J!(+#$0&0)56'&!,+b#+&'+J!'(+95'J!$+,M5$'20$N!*5,2$+,,J!7NM0)&N6+95'J!'(*!609M&56'250(,!08!M$+)('(6Nz!K9!=![$0M!B+*!UN)J!US\!ZI>`IE!
B#27',J!/EJ!O';(5,J!jE!KEJ!V#(*;0$)J!BEJ!'(*!W'$&+(J!KEJ!CDDFJ!X,!52!M0,,5;&+!20!5(6$+',+!752!$'2+,!5(!,2$#62#$+>;',+*!@5$2#'&!,6$++(5()!;N!M7'$9'60M70$+!85&2+$5()é!K(!5(@+,25)'250(!08!27+!'*@'(2')+,!'(*!M528'&&,!08!M0,2>85&2+$5()z!=!B0&!-$'M7E!B0*+&J!JU\!GCYI>GCZGE!
3'#&'J!cEJ!L'$,0(,J!BEJ!'(*!B022$'9J!=E!cEJ!CDDZJ!L$02+5(!15(',+,!',!*$#)!2'$)+2,!5(!2$NM'(0>,09+,!'(*!V+5,79'(5'z!<506759E!<50M7N,E!K62'J!HcTS\!GZG>GZHE!
3++*&+9'(J!OE!<E!'(*!_#(,67J!cE!/EJ!GHIDJ!K!)+(+$'&!9+270*!'MM&56';&+!20!27+!,+'$67!80$!,595&'$525+,!5(!27+!'95(0!'65*!,+b#+(6+!08!240!M$02+5(,z!=!B0&!<50&J!SR\!]]Y>]ZYE!
3+&,0(J!/E!VE!'(*!c0\J!BE!BEJ!CDDGJ!V+7(5()+$!>!<5067+95+Ez!OM$5()+$!R+$&')J!<+$&5(J!U+5>*+&;+$)J!3+4!j0$1E!
3+$@'&&J!BEJ!U'(,M+$,J!LEJ!c'$&,,0(J!=EJ!<0#17'$2'J!VEJ!'(*!Kb@5,2J!=EJ!CDDFJ!L$+*5625()!;5(*5()!90*+,!8$09!8$++!+(+$)N!6'&6#&'250(,z!=!B+*!c7+9J!TH\!C`ZI>C``IE!
35670&',J!WE!<EJ!35670&',J!UE!<EJ!'(*!/++$85+&*J!/E!_EE!-+(+/06z!K('&N,5,!'(*!R5,#'&5"'250(!08!-+(+256!R'$5'250(E!%9;(+4E!3+4,E!GHHIE!!
!356#&+,6#>/#@'"J!/EJ!_7522'1+$J!OEJ!OM$5()+$J!cEJ!'(*!B'$'5,J!PEJ!CDDIJ![7+!%-:!$+6+M20$!
U01+N>c01+Nz!c'(6+$!c+&&J!HH\!CDH>CGGE!
35,,5(1J!=E!_EJ!B#$$'NJ!cEJ!U'$2,70$(J!BEJ!R+$*0(1J!BE!VEJ!c0&+J!=E!cEJ!'(*!['N&0$J!PEJ!CDDCJ!K!(+4!2+,2!,+2!80$!@'&5*'25()!M$+*56250(,!08!M$02+5(>&5)'(*!5(2+$'6250(z!L$02+5(,J!Se\!]ZI>]IGE!
30;&+J!BE!%EJ!%(*56022J!=E!KEJ!'(*!=07(,0(J!VE!3EJ!CDD]J!L$02+5(!15(',+!5(75;520$,z!5(,5)72,!5(20!*$#)!*+,5)(!8$09!,2$#62#$+z!O65+(6+J!POP\!GFDD>GFDZE!
LITERATURVERZEICHNIS
GGF!
30$0J!PE!'(*!-+99'J!KEJ!CDDFJ!%-:P!'(*!)+8525(5;!eX$+,,'fz!-'(![0!W')'1#!PN070J!PT\!GDI`>GDIHE!
34'1'J!OE!'(*!P5*&+NJ!PE!-EJ!CDDYJ!R5$2#'&!*$#)!*5,60@+$N!'(*!*+@+&0M9+(2!80$!(+)&+62+*!*5,+',+,!27$0#)7!M#;&56>M$5@'2+!M'$2(+$,75M,z!3'2!P+@!/$#)!/5,60@J!J\!HGH>HCFE!
d+&&5+(J!:EJ!c$'9+$J!=EJ!<+N+$J!cEJ!X7&+(8+&*2J!_E!/EJ!'(*!O+&"+$J!LE!BEJ!CDD`J![7+!59M'62!08!2'#209+$!80$9,!0(!M7'$9'60M70$+>;',+*!@5$2#'&!,6$++(5()z!=!c7+9!X(8!B0*+&J!SU\!CY]C>CYZ]E!
d+&&5+(J!:EJ!%()+&,J!WEJ!c$'9+$J!=EJ!B'$7g8+$J!PE!=EJ!W+$(J!cEJ!'(*!O+&"+$J!LE!BEJ!CDDHJ!O+'$67>5()!(+4!K(25M'$',5256,!5(!R5$2#'&!OM'6+!in!:$09!B0&+6#&'$!['$)+2,!20!/$#)!c'(*5*'2+,z!O+&"+$J!LE!BEJ!%*EJ!_XV%j>RcU!R+$&')!-9;U!k!c0E!W-'KJ!_+5(7+59E!
d+&&5+(J!:EJ!X7&+(8+&*2J!_E!/EJ!'(*!-',2+5)+$J!=EJ!CDDZJ!X(8R5,>>M&'280$9>5(*+M+(*+(2!@5,#'&!*'2'!95(5()!08!9#&25*59+(,50('&!67+956'&!*'2'!,+2,z!=!c7+9!X(8!B0*+&J!ST\!G]Z`>G]`IE!
d(0*+$'J!WEJ!O'20#J!WEJ!'(*!U5$02'J!UEJ!CDDIJ!%@'&#'250(,!08!90&+6#&'$!*0615()!M$0)$'9,!80$!@5$2#'&!,6$++(5()z!=!c7+9!X(8!B0*+&J!Sc\!G`DH>G`GFE!
d$"'+"J!BEJ!-0$2'2J!KEJ!B0(*$')0(J!VEJ!<'67,J!dEJ!'(*!L+$+">L'N'J!%EJ!CDDHJ!K[L>(0(609M+2525@+!5(75;520$,!08!c/W>6N6&5(!609M&+\+,z!c7+9B+*c7+9J!S\!GH>C]E!
d,&0;J!=E!/EJ!U+#9'((J!OE!KEJ!j#J!cE!UEJ!K&&+(J!/E!KEJ!<',1'$'(J!OEJ!<#5J!BEJ!/+&'$0,'J!%EJ!:#()J!KE!/EJ!U',7',7J!KEJ!U'#J!=EJ!X@NJ!OEJ!='60;,J!=E!_EJ!V+4J!_EJ!B'#()J!=EJ!B6/04>+&&J!PE!OEJ!P52675+J!OEJ!P09'(04,15J!BE!=EJ!O5&@+$9'(J!=E!KEJ!j'()J!_EJ!^70()J!BEJ!'(*!:#67,>W(022,J![EJ!CDDHJ!_'2+$>,0&#;&+!M$0*$#),!08!'(!K#$0$'!15(',+!5(75;520$z!<500$)E!B+*!c7+9!V+22J!He\!G]DH>G]GCE!
L'@&+2567J!3E!LEJ!GHHHJ!B+67'(5,9,!08!6N6&5(>*+M+(*+(2!15(',+!$+)#&'250(z!,2$#62#$+,!08!c*1,J!27+5$!6N6&5(!'625@'20$,J!'(*!c5M!'(*!X3W]!5(75;520$,z!=!B0&!<50&J!JRc\!FCG>FCFE!
L'N(+J!BE!cEJ![+2+$J!BE!LEJ!K&&'(J!/E!cEJ!K$5',J![E!KEJ!'(*!=0'((0M0#&0,J!=E!/EJ!GHHCJ!X2+$'25@+!95(595"'250(!2+67(5b#+,!80$!';!5(5250!202'&>+(+$)N!6'&6#&'250(,z!B0&+6#&'$!*N('956,!'(*!60(p#)'2+!)$'*5+(2,z!P+@5+4,!08!B0*+$(!L7N,56,J!US\!GD]Z>GDHIE!
L+$0&'J!%EJ!CDD`J!B5(595"5()!8'&,+!M0,525@+,!5(!15(',+!@5$2#'&!,6$++(,z!L$02+5(,J!US\!]CC>]YZE!
L$'22J!/E!=EJ!<+(2&+NJ!=EJ!=+4,;#$NJ!LEJ!<0N&+J!:E![EJ!%(*56022J!=E!KEJ!'(*!30;&+J!BE!%EJ!CDD`J!/5,,+625()!27+!*+2+$95('(2,!08!6N6&5(>*+M+(*+(2!15(',+!C!'(*!6N6&5(>*+M+(*+(2!15(',+!]!5(75;520$!,+&+625@52Nz!=!B+*!c7+9J!Se\!Z]ID>Z]IIE!
i#J!}EJ!O4'(,0(J!PEJ!/'NJ!PEJ!'(*![,'5J!=EJ!CDDHJ!K!)#5*+!20!2+9M&'2+!;',+*!,2$#62#$+!M$+*56>250(z!c#$$!L$02+5(!L+M2E!O65J!HO\!CID>CFZE!
i#5(2',>c'$*'9'J!KE!'(*!c0$2+,J!=EJ!CDDIJ!35&025(5;!27+$'MN!5(!67$0(56!9N+&0)+(0#,!&+#1+>95'z!/$#),![0*'N!e<'$6E!fJ!SP\!`HG>IDCE!
i#5(2#,J!:EJ!OM+$'(*50J!dEJ!-$N(;+$)J!=EJ!L+252p+'(J!BEJ!'(*![#88+$NJ!LEJ!CDDHJ!V5)'(*!,6'880&*!70MM5()!609;5(5()!Y/!9'\59'&!,#;,2$#62#$+!,+'$67!'(*!90&+6#&'$!,595&'$52Nz!<Bc!<505(80$9'256,J!HO\!C]ZE!
! ! LITERATURVERZEICHNIS
GGH!
P'9'67'(*$'(J!-E!3EJ!P'9'1$5,7('(J!cEJ!'(*!O',5,+17'$'(J!REJ!GH`YJ!O2+$+067+95,2$N!08!M0&NM+M25*+!67'5(!60(85)#$'250(,z!=!B0&!<50&J!c\!HZ>HHE!
P'N90(*J!=E!_EJ!_'2,0(J!XE!KEJ!'(*!B'70#5J!KEJ!CDDHJ!P'250('&5"5()!&+'*!0M2595"'250(!;N!',,065'25()!b#'(252'25@+!$+&+@'(6+!4527!90&+6#&'$!,2$#62#$+!90*5856'250(z!=!c7+9!X(8!B0*+&J!Se\!GHZC>GH`CE!
P+)'""5J!BE!<EJ!R5&&'(5J!LEJ!B',+$'25J!PEJ!c06675J!VEJ!-5'6675(0J!PEJ!<#$$0(5J!/EJ!'(*!P+22'(5J!BEJ!GHHHJ!L7'$9'6015(+256!@'$5';5&52N!'(*!,2$'2+)N!80$!27+$'M+#256!*$#)!90(520$5()!08!,'b#5('@5$!eOiRf!5(!UXR>G!5(8+62+*!5(*5@5*#'&,z!<$!=!c&5(!L7'$9'60&J!Sc\!YIH>YFCE!
P0)+$,J!/EJ!<$04(J!PE!/EJ!'(*!U'7(J!BEJ!CDDZJ!?,5()!+\2+(*+*>60((+625@52N!85()+$M$5(2,!4527!V'M&'65'(>90*585+*!<'N+,5'(!'('&N,5,!5(!75)7>27$0#)7M#2!,6$++(5()!80&&04>#Mz!=!<50>90&E!O6$++(EJ!HO\!`FC>`F`E!
P00,J!/E!OEJ!c$'480$*J!BE!=EJ!/0('&*J!PE!-EJ!:$'#(70&"J!BEJ!U'$;J!dE!OEJ!U+J!cE!jEJ!W5,,5()+$J!=E!cEJ!O7'4J!BE!WEJ!'(*!O2$5+M+(J!<EJ!CDDCJ!B5(5()!27+!L&',90*5#9!)+(09+!*'2';',+!20!*+85(+!0$)'(+&&'$!8#(6250(z!47'2!*0+,!27+!'M560M&',2!*0éz!L75&0,E![$'(,!P!O06!V0(*!<!<50&!O65J!PTc\!YZ>]`E!
P0,,>B'6*0('&*J!LE!<EJ!-$'+,+$J!PEJ!W'MM+,J!<EJ!:$'(1&5(J!PEJ!'(*!_5&&5'9,0(J!/E!UEJ!GHH]J!X,0&'250(!'(*!+\M$+,,50(!08!'!)+(+!,M+658N5()!'!6*6C>&51+!M$02+5(!15(',+!8$09!27+!7#9'(!9'&'$5'!M'$',52+!L&',90*5#9!8'&65M'$#9z!%#$!=!<5067+9J!JJO\!`HY>IDGE!
O'*04,15J!=EJ!GHHIJ!K!7N;$5*!'MM$0'67!80$!'**$+,,5()!$5()!8&+\5;5&52N!5(!Y/!*'2';',+!,+'$67>5()z!=!c09M#2E!K5*+*!B0&!/+,J!HH\!ZY>`DE!
O'2N'('$'N'('J!KE!'(*!W'&*5,J!LEJ!CDDHJ!B'99'&5'(!6+&&>6N6&+!$+)#&'250(z!,+@+$'&!c*1,J!(#>9+$0#,!6N6&5(,!'(*!*5@+$,+!609M+(,'20$N!9+67'(5,9,z!d(60)+(+J!JR\!CHCZ>CHYHE!
O67++(J!KE!=E!'(*!L'b#02J!3EJ!CDDHJ!R5&*')&5M25(!e-'&@#,f!'(*!85\+*!609;5('250(!@5&>*')&5M25(+>9+280$95(!e%#6$+',f!5(!27+!2$+'29+(2!08!2NM+!C!*5';+2+,z!P+@!B+*!V5+)+J!US\!G`G>G`IE!
O67&561J![E!'(*!:0)+&,0(J!KEJ!GHHCJ![3LKcW!>>!K!2$#(6'2+*!3+420(!95(595"'250(!M'61')+!80$!&'$)+>,6'&+!M$0;&+9,Ez!KcBE![$'(,E!B'27E!O0824'$+J!HR\!]`>GGGE!
O67(+61+J!RE!'(*!<0,2$09J!=EJ!CDD`J!c09M#2'250('&!67+95,2$N>*$5@+(!*+65,50(!9'15()!5(!&+'*!)+(+$'250(z!/$#)!/5,60@![0*'NJ!HH\!]Y>ZDE!
O67(+5*+$J!-E!'(*!:+67(+$J!?EJ!CDDZJ!c09M#2+$>;',+*!*+!(0@0!*+,5)(!08!*$#)>&51+!90&+6#&+,z!3'2!P+@!/$#)!/5,60@J!S\!`]H>``YE!
O674'$*2J!dEJ!W0&;J!UEJ!'(*!%$(,2J!<EJ!CDDYJ!/$#)!*5,60@+$N!20*'Nz!c#$$![0M!B+*!c7+9J!P\!G>HE!
O6&'8'(5J!PE!KEJ!GHH`J!cN6&5(!*+M+(*+(2!15(',+!'625@'25()!15(',+,z!c#$$!dM5(!c+&&!<50&J!R\!IFF>IH]E!
LITERATURVERZEICHNIS
GCD!
O+&"+$J!LE!BEJ!CDDZJ!O2$#62#$+>;',+*!$'250('&!*$#)!*+,5)(z!3+#+!Wege!*+$!90*+$(+(!_5$1>,2088>!+(24561&#()E!X(z!V#65#,J!PEn!U5+M+J![En!-022,2+5(J!<E!eU$,)f!-$#(*"Q)+!*+$!'&&)+>9+5(+(!L'$',520&0)5+z!L'$+NJ!<+$&5(E!
O+&"+$J!LE!BEJ!B'$708+$J!PE!=EJ!'(*!P074+$J!KEJ!CDDFJ!KMM&5+*!<505(80$9'256,z!K(!X(2$0*#6>250(Ez!OM$5()+$!R+$&')J!<+$&5(J!U+5*+&;+$)J!3+4!j0$1E!
O7+5(+$9'(J!:E!<EJ!-5$'#*J!%EJ!'(*!V'0#5J!KEJ!CDDZJ!U5)7!'885(52N!2'$)+2,!08!M$02+5(!15(',+!5(75;520$,!7'@+!,595&'$!$+,5*#+,!'2!27+!M0,5250(,!+(+$)+256'&&N!59M0$2'(2!80$!;5(*5()z!=!B0&!<50&J!PTJ\!GGY]>GGZ`E!
O75$&+NJ!BE!_EJ!GHFHJ!/+@+&0M9+(2!08!'!&5@+!'22+(#'2+*!@'665(+!')'5(,2!60665*50,5,!08!M0#&>2$Nz!L'$',52+!X99#(0&J!HH\!GGI>GC]E!
O75$&+NJ!BE!_EJ!O9527J!KE!VEJ!'(*!<&'1+J!/E!LEJ!CDDIJ!c7'&&+()+,!5(!27+!,#66+,,8#&!60(2$0&!08!27+!'@5'(!60665*5'z!R'665(+J!JT\!ZZ]D>ZZ]IE!
O0(5J!PEJ!dâP+5&&NJ![EJ!:#$+2J!LEJ!B#&&+$J!VEJ!O2+M7'(J!cEJ!^#9,2+5(>B+61+$J!OEJ!:$+2"J!UEJ!:';>;$0J!/EJ!'(*!c7'#*7#$5J!<EJ!CDDGJ!O+&+625@+!5(!@5@0!'(*!5(!@52$0!+88+62,!08!'!,9'&&!90&+>6#&+!5(75;520$!08!6N6&5(>*+M+(*+(2!15(',+!]z!=!3'2&!c'(6+$!X(,2J!eP\!]Y`>]]`E!
O02$588+$J!cE!KE!'(*!/$'9;#$)J!XEJ!CDDZJ!.X(!,52#!6$0,,>*0615().!20!,59#&2'(+0#,&N!'**$+,,!9#&25M&+!2'$)+2,z!=!B+*!c7+9J!SR\!YGCC>YGCZE!
O0#,'J!OE!:EJ!:+$('(*+,J!LE!KEJ!'(*!P'90,J!BE!=EJ!CDD`J!L$02+5(>&5)'(*!*0615()z!6#$$+(2!,2'2#,!'(*!8#2#$+!67'&&+()+,z!L$02+5(,J!UT\!GZ>C`E!
O2'7&J!BE!KEJ!CDDDJ!O2$#62#$+><',+*!V5;$'$N!/+,5)(J!5(!Virtual Screening for Bioacitve Mole-culesz!_5&+N>RcU!R+$&')J!_+5(7+59J!CCH>C`]E!
O2+M7+(J!<EJ!P099+&J!BEJ!/'#),675+,J!KEJ!'(*!U';+$10$(J!KEJ!GHHIJ!O2#*5+,!08!$+,5,2'(6+!20!'(2560665*5'&,!5(!%59+$5'!85+&*!5,0&'2+,!'(*!M#$+!%59+$5'!,2$'5(,z!R+2!L'$',520&J!Ue\!GH>CHE!
O2p+$(,67'(2"J!%EJ!B'$+&5#,J!=EJ!B+*5('J!cEJ!='60;,,0(J!BEJ!R+$9+#&+(J!3E!LEJ!'(*!d0,2+(>;$5(1J!cEJ!CDD`J!K$+!'#209'2+*!90&+6#&'$!*N('956,!,59#&'250(,!'(*!;5(*5()!8$++!+(+$)N!6'&6#&'250(,!$+'&5,256!200&,!5(!&+'*!0M2595"'250(é!K(!+@'&#'250(!08!27+!&5(+'$!5(2+$'6250(!+(+$)N!eVX%f!9+270*z!=!c7+9!X(8!B0*+&J!SU\!GHIC>GHFYE!
['82J!cE!KEJ!/'!O5&@'J!RE!<EJ!'(*!/'!O5&@'J!cE!UEJ!CDDFJ!c#$$+(2!20M56,!5(!609M#2+$>'5*+*!*$#)!*+,5)(z!=!L7'$9!O65J!ec\!GDFH>GDHFE!
[+210J!XE!REJ!CDDZJ!c09M#25()!67+95,2$N!0(!27+!4+;z!/$#)!/5,60@![0*'NJ!HO\!G]HI>GZDDE!
[709M,0(J!=E!/EJ!U5))5(,J!/E!-EJ!'(*!-5;,0(J![E!=EJ!GHH]J!cV?O[KV!_z!59M$0@5()!27+!,+(,5>25@52N!08!M$0)$+,,5@+!9#&25M&+!,+b#+(6+!'&5)(9+(2!27$0#)7!,+b#+(6+!4+5)725()J!M0,5250(>,M+65856!)'M!M+('&25+,!'(*!4+5)72!9'2$5\!67056+z!3#6&+56!K65*,!P+,J!JJ\!]`IY>]`FDE!
[095(')'J!jE!'(*!=0$)+(,+(J!_E!VEJ!CDD]J!-+(+$'&!90*+&!80$!+,259'250(!08!27+!5(75;5250(!08!M$02+5(!15(',+,!#,5()!B0(2+!c'$&0!,59#&'250(,z!=!B+*!c7+9J!Sc\!CZY]>CZ]HE!
! ! LITERATURVERZEICHNIS
GCG!
[$'\&+$J!LEJ!-$++(J!=EJ!B+22J!UEJ!O+b#5(J!?EJ!'(*!:#$+2J!LEJ!GHHHJ!?,+!08!'!M7'$9'60M70$+!90*+&!80$!27+!*+,5)(!08!%-:P!2N$0,5(+!15(',+!5(75;520$,z!5,08&'@0(+,!'(*!Y>M7+(N&>]eGUf>b#5(0&0(+,z!=!B+*!c7+9J!SJ\!GDGF>GDC`E!
_'&&+$J!PE!:EJ!W++&5()J!LE!=EJ!/0('&*J!PE!-EJ!O2$5+M+(J!<EJ!U'(*9'(J!%EJ!V'()>?((',67J!3EJ!c049'(J!KE!:EJ!<+,$'J!-E!OEJ!P00,J!/E!OEJ!'(*!B6:'**+(J!-E!XEJ!GHHFJ!3#6&+'$>+(60*+*!M$02+5(,!2'$)+2!20!27+!M&',25*!5(![0\0M&',9'!)0(*55!'(*!L&',90*5#9!8'&65M'$#9z!L$06!3'2&!K6'*!O65!?!O!KJ!eT\!GCYZC>GCYZIE!
_'&&+$J!PE!:EJ!P++*J!BE!<EJ!c049'(J!KE!:EJ!'(*!B6:'**+(J!-E!XEJ!CDDDJ!L$02+5(!2$'885615()!20!27+!M&',25*!08!L&',90*5#9!8'&65M'$#9!5,!@5'!27+!,+6$+20$N!M'274'Nz!%B<d!=J!He\!GIH]>GFDCE!
_'&2+$,J!_E!LEJ!O2'7&J!BE!KEJ!'(*!B#$610J!BE!KEJ!GHHFJ!R5$2#'&!O6$++(5()!>!K(!d@+$@5+4z!/$#)!/5,60@E![0*'NJ!P\!G`D>GIFE!
_'$*J!LEJ!%b#5(+2J!VEJ!L'61+$J!=EJ!'(*!/0+$5)J!cEJ!CDD]J!L$02+5(!15(',+,!08!27+!7#9'(!9'&'$5'!M'$',52+!L&',90*5#9!8'&65M'$#9z!27+!15(09+!08!'!*5@+$)+(2!+#1'$N02+z!<BcE!-+(09>56,J!T\!IHE!
_'$$+(J!-E!VEJ!K(*$+4,J!cE!_EJ!c'M+&&5J!KE!BEJ!c&'$1+J!<EJ!V'V0(*+J!=EJ!V'9;+$2J!BE!UEJ!V5(*@'&&J!BEJ!3+@5(,J!3EJ!O+9#,J!OE!:EJ!O+()+$J!OEJ![+*+,60J!-EJ!_'&&J!XE!/EJ!_00&@+(J!=E!BEJ!L+5,7088J!cE!%EJ!'(*!U+'*J!BE!OEJ!CDD`J!K!6$5256'&!',,+,,9+(2!08!*0615()!M$0)$'9,!'(*!,60$5()!8#(6250(,z!=!B+*!c7+9J!Se\!ZHGC>ZHYGE!
_'2+$,J!3E!cEJ!_00*'$*J!cE!VEJ!'(*!L$5))+J!OE![EJ!CDDDJ!cN6&5(!U!'625@'250(!'(*!*$#)!,#,6+M>25;5&52N!08!27+!L89$1!6N6&5(!*+M+(*+(2!M$02+5(!15(',+!8$09!L&',90*5#9!8'&65M'$#9z!B0&!<5067+9!L'$',520&J!HOc\!]Z>ZZE!
_7++&+$J!/E!VEJ!c7#$67J!/E!BEJ!%*)'$J!PEJ!:+*+$7+(J!OEJ!U+&9;+$)J!_EJ!B'**+(J![E!VEJ!L0(>25#,J!=E!?EJ!O67#&+$J!-E!/EJ!O67$59&J!VE!BEJ!O+b#+5$'J!%EJ!O#"+1J![E!dEJ!['2#,0@'J![E!KEJ!'(*!_')(+$J!VEJ!CDD]J!/'2';',+!$+,0#$6+,!08!27+!3'250('&!c+(2+$!80$!<502+67(0&0)N!X(>80$9'250(z!#M*'2+z!3#6&+56!K65*,!P+,J!PJ\!/YZ>/]DE!
_5&&+22J!LEJ!CDDYJ!O595&'$52N>;',+*!'MM$0'67+,!20!@5$2#'&!,6$++(5()z!<5067+9!O06![$'(,J!PH\!`DY>`D`E!
_5&&+22J!LEJ!CDD`J!O595&'$52N>;',+*!@5$2#'&!,6$++(5()!#,5()!C/!85()+$M$5(2,z!/$#)!/5,60@![0>*'NJ!HH\!GD]`>GDZYE!
_5&&5'9,J!PE!<EJ!c'$&N&+J!_E!_EJ!<0(*J!/E!PEJ!'(*!<$04(J!XE!KEJ!GHHHJ![7+!+8856'6N!'(*!+60>(0956!;+(+852,!08!L'$'60\J!'!&5@+!'22+(#'2+*!'(2560665*5'&!@'665(+J!5(!6099+$65'&!2$5'&,!4527!,2'(*'$*!;$05&+$!67561+(,!5(!27+!?(52+*!W5()*09z!X(2!=!L'$',520&J!Je\!Y]G>YZZE!
_5&,0(J!PE!=EJ!/+((NJ!LE!_EJ!L$+5,+$J!LE!PEJ!P'()'67'$5J!WEJ!P0;+$2,J!WEJ!P0NJ!KEJ!_7N2+J!KEJ!O2$'27J!BEJ!B00$+J!/E!=EJ!B00$+J!LE!_EJ!'(*!_5&&5'9,0(J!/E!UEJ!GHH`J!c09M&+2+!)+(+!9'M!08!27+!M&',25*>&51+!/3K!08!27+!9'&'$5'!M'$',52+!L&',90*5#9!8'&65M'$#9z!=!B0&!<50&J!JUH\!GZZ>GICE!
_5,,&+$J!cEJ!GHDZJ
LITERATURVERZEICHNIS
GCC!
_00*'$*J!cE!VEJ!V5J!^EJ!W'276'$2J!KE!WEJ![+$$+&&J!=EJ!-+$+('J!VEJ!V0M+">O'(67+"J!BEJ!WN&+J!/E!%EJ!<7'22'67'$p++J!KE!WEJ!35670&,J!/E!KEJ!%&&5,J!_EJ!L$5))+J!OE![EJ!-+N+$J!=E!KEJ!'(*!_'>2+$,J!3E!cEJ!CDDYJ!d\5(*0&+>;',+*!609M0#(*,!'$+!,+&+625@+!5(75;520$,!08!L&',90*5#9!8'&65M'$#9!6N6&5(!*+M+(*+(2!M$02+5(!15(',+,z!=!B+*!c7+9J!SU\!YFII>YFFCE!
_#J!OE!jEJ!B63'+J!XEJ!W0(20M5*5,J!-EJ!B6c&#+J!OE!=EJ!B6X((+,J!cEJ!O2+4'$2J!WE!=EJ!_'()J!OEJ!^7+&+@'J!/E!XEJ!B'$$5')+J!UEJ!V'(+J!/E!LEJ!['N&0$J!LEJ!:5,67+$J!LE!BEJ!'(*!_'&15(,7'4J!BE!/EJ!CDDYJ!/5,60@+$N!08!'!(0@+&!8'95&N!08!c/W!5(75;520$,!4527!27+!M$0)$'9!VX>/K%?Oz!,2$#62#$'&!;',5,!80$!&5)'(*>5(*#6+*!*5,0$*+$5()!08!27+!'625@'250(!&00Mz!O2$#6>2#$+J!HH\!YHH>]GDE!
}5'()J!^EJ!CDD`J!K*@'(6+,!5(!7090&0)N!M$02+5(!,2$#62#$+!90*+&5()z!c#$$!L$02+5(!L+M2E!O65J!c\!CGI>CCIE!
j00(J!OE!'(*!_+&,7J!_E!=EJ!CDD]J!X*+(25856'250(!08!'!95(59'&!,#;,+2!08!$+6+M20$!60(80$9'250(,!80$!59M$0@+*!9#&25M&+!60(80$9'250(!*0615()!'(*!240>,2+M!,60$5()z!=!c7+9!X(8!c09M#2E!O65J!SS\!FF>H`E!
j#()J!OEJ!?((',67J![E!PEJ!'(*!V'()>?((',67J!3EJ!CDDGJ!K('&N,5,!08!'M560M&',2!2'$)+25()!'(*!2$'(,52!M+M25*+!M$06+,,5()!5(![0\0M&',9'!)0(*55!;N!*+&+250('&!'(*!5(,+$250('&!9#2')+(+>,5,z!B0&!<5067+9!L'$',520&J!HHR\!GG>CGE!
^7'5J!OEJ!O+(*+$0456"J!KE!BEJ!O'#,@5&&+J!%E!KEJ!'(*!:5))J!_E!/EJ!CDDCJ!:&'@0M5$5*0&J!'!(0@+&!6N6&5(>*+M+(*+(2!15(',+!5(75;520$J!5(!6&5(56'&!*+@+&0M9+(2z!K((!L7'$9'6027+$EJ!PU\!HDZ>HGGE!
^#+))+J!=EJ!P'&M7J!OEJ!O679#1+$J!BEJ!B6:'**+(J!-E!XEJ!'(*!O67(+5*+$J!-EJ!CDDGJ!/+65M7+$5()!'M560M&',2!2'$)+25()!,5)('&,>>8+'2#$+!+\2$'6250(!8$09!(#6&+'$>+(60*+*!M$+6#$,0$,!08!L&',90*5#9!8'&65M'$#9!'M560M&',2!M$02+5(,z!-+(+J!JRO\!GH>C`E!
!!