Sentetik maddeler ile DNA kesimi

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1. 4. 3. 1. Antrasiklin Antitümör Antibiyotikleri 14 1. 4. 3. 2. Bleomycin 15 1. 4. 3. 3. Enediyen Antibiyotikleri 16 1. 4. 3. 3. 1. Neocarzinostatin 16 1. 4. 3. 3. 2. Esperamicin ve Calicheamicin 18 1. 4. 3. 3. 3. Dynemicin A 20 1. 5. Karbon Merkezli Radikal Metabolitleri 20

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.XOODQÕODQ$OHWOHU 36 3. 2. Metot 36 3. 2. 1. Agaroz Jel Elektroforezi 36

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AMAÇ

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ÖZET

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$\UÕFD DULO UDGLNDOHUL LOH ROXúDQ '1$ KDVDUÕ ]Hrine antioksidant ve antimikrobiyal aktiviteleri bilinen Hypericum retusum Aucher, Hypericum scabrum L.,

Hypericum lysimachioides Boiss&Nöe var .lysimachioides, Achillea aleppica D.C. subsp. aleppica, Achillea aleppica D.C. subsp. zederbaueri (Hayek) Hub. -Mor. ve

Achillea biebersteinii $IDQ ELWNL HNVWDUDNODUÕQÕQ NRUX\XFX HWNLOHUL DJDUR] MHO

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ELWNLOHULQ LoLQGH EXODQDQ IODYRQRLGOHULQ EDNÕU LOH úHODWODúDUDN JHUoHNOHúWL÷L GúQOPHNWHGLU

Anahtar Kelimeler: '1$ .HVLPL 6HUEHVW 5DGLNDOOHU 5DGLNDO 6|QGUF %DNÕU

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ABSTRACT

The ultimate aim of this thesis was to investigate the effect of different aryl substituted analogues of benzenediazonium tetrafluoroborate on radical reactivity and DNA cleavage ability. The results indicated that cleavage of a DNA duplex from pBluescript M13+ DNA was observed by the different substituted aryl diazonium salts, in the presence of inorganic one-electron donor. Moreover, DNA cleavage was partly affected by different substitute group and DNA strand-scission was also observed without one electron donor in the presence of some tested diazonium salts. Its mechanism involves the generation of the aryl radical that cleaving DNA by hydrogen atom abstraction from deoxyribose sugar.

The protective effect of Hypericum retusum Aucher, Hypericum scabrum L.,

Hypericum lysimachioides Boiss&Nöe var .lysimachioides, Achillea aleppica D.C. subsp. aleppica, Achillea aleppica D.C. subsp. zederbaueri (Hayek) Hub. -Mor. ve

Achillea biebersteinii Afan. plants extracts on DNA cleavage mediated by aryl radical was also investigated using pBluescript M13+ plasmid DNA, as judged by agarose gel electrophoresis. It was found that aryl radical induced DNA strand scission was not prevented by ethanol extracts of Hypericum and Achillea. On the contrary, DNA cleavage was increased in the presence of plant extracts. Therefore, we decided to investigate the DNA cleavage ability of plant extracts alone and also in the presence of copper (II) chloride. It was found that the naturally occuring plant extracts caused strand scission of DNA in the presence of copper ion. The mechanism of the DNA strand scission by plant extracts (in the presence of Cu2+ ion) indicated an involvement of oxygenation of aromatic nucleus, affording catecholic moieties that were proposed to coordinate Cu2+ and subsequently effect the reduction of dioxygen to reactive species, followed by the oxidation of catecholic moiety via the coordinated Cu2+ ion.

Keywords: DNA Cleavage, Free Radical, Radical Scavenger, Copper, Hypericum,

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KISALTMALAR

DNA Deoksiribonükleik asit RNA Ribonükleik asit

A1 Achillea aleppica subsp. aleppica A2 Achillea aleppica subsp. zederbaueri A3 Achillea biebersteinii

H1 Hypericum retusum

H2 Hypericum scabrum H3 Hypericum lysimachioides

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1. 2. DNA’ya Zarar Veren Maddeler Neden Önemlidir?

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RODQ GR÷DO UQOHU ED]HQ EHNOHQPHGLN \ROODUOD EL\RORMLN HWNL J|VWHUHELOGLNOHUL LoLQ NRPSOHNV\DúDPVUHFLQLDoÕNODPDNLoLQ\DUDUOÕRODELOLUOHU

.RYDOHQW'1$0RGLILNDV\RQXQXQ*HQHO0HNDQL]PDVÕ

'1$¶QÕQNRYDOHQWPRGLILNDV\RQPHNDQL]PDODUÕLNLNDWHJRULGHVÕQÕIODQGÕUÕOÕU(i) (OHNWURILOOHULQ'1$¶QÕQQNOHRILOLNNÕVÕPODUODUHDNVL\RQX\DGDLLUDGLNDOOHULQ'1$ ile reaksiyonu.

(OHNWURILOOHU '1$ \DSÕVÕQGDNL IDUNOÕ QNOHRILOLN NÕVÕPODUOD HWNLOHúHELOLU .LP\DVDO \DSÕODUÕQD ED÷OÕ RODUDN '1$¶QÕQ EHOOL QNOHRILOLN NÕVÕPODUÕQD VHOHNWLYLWH gösterirler. DNA’da guaninin N-7, N-3 ve exosiklik N2 amino grubu ile adeninin N-7 ve N-NÕVÕPODUÕGR÷DOUQOHUOHHQoRNPRGLILNDV\RQDX÷UD\DQNÕVÕPODUGÕU17-20_ùHNLO

(17)

ùHNLOWatson–&ULFNED]oLIWLYHúHNHUIRVIDWRPXUJDVÕ ùHNLO$ED]LNE|OJHROXúXPXYH'1$NHVLPL O N _N N NH O NH₂ RO RO O N _N N NH O NH₂ RO RO E E+ + N _N N NH O NH₂ E O RO RO H2 O O RO RO OH OH RO RO O OH RO RO O + (1) + N N O O CH₃ H R N _N N N N H H R N N O N H H R N _N N N O N H H H R 1 2 3 5 6 7 3 8 9 Büyük Oluk Küçük Oluk A-T G-C Büyük Oluk Küçük Oluk 5 3 1 2 3 1 6 8 9 R=DNA Zinciri O B O P O O O O H O B O H -B=DNA Baz 3' C5 ' C4' C3' C2 ' C1'

(18)

3ULQ NDOÕQWÕODUÕQÕQ 1-7 ya da N-3 pozisyonundan elektrofilik modifikasyonu JOLNR]LGLNED÷ÕQ]D\ÕIODPDVÕ\ODVRQXoODQÕU%XQXQVRQXQGDDED]LNE|OJH1ROXúXUYH Q|WUDO úDUWODUGD EX E|OJHOHU KLGUROL]OHQHUHN '1$ NHVLPL JHUoHNOHúLU21-25_ùHNLO

'1$ ED]ODUÕQGDNL H[RVLNOLN D]RW YH NDUERQLO RNVLMHQOHUL \D GD '1$ RPXUJDVÕQGDNL IRVIDW RNVLMHQOHUL LOH HOHNWURILN WUOHUL UHDNVL\RQX JHQHOOLNOH NDUDUOÕ UQOHU ROXúWXUXU26-28

%D]Õ UDGLNDO WUOHUL '1$ LOH GH÷LúLN SR]LV\RQODUGD HWNLOHúLUOHU '1$¶QÕQ UDGLNDOOHU WDUDIÕQGDQ KDVDUD X÷UDWÕOPDVÕQGD HQ |QHPOL \RO GHoksiribozdan hidrojen DWRPX NRSDUÕOPDVÕGÕU29-31 _{'HRNVLULER] úHNHULQGHQ KLGURMHQ DWRPX NRSDUÕOPDVÕ úHNHU}

IRVIDW RPXUJDVÕQÕQ NÕUÕOPDVÕQD VHEHS ROXU30-31_{ùHNLO ùHNHU IRVIDW RPXUJDVÕQD}

HWNLPH\HHNRODUDNED]ÕUDGLNDOOHUGH'1$ED]ODUÕ\ODHWNLOHúLUOHU32-35_{Bu reaksiyonlar}

úHNHU IRVIDW RPXUJDVÕQÕQ NÕUÕOPDVÕQD VHEHS ROPD]ODU IDNDW '1$ \DSÕVÕQGDNL PRGLILNDV\RQODUEL\RORMLNRODUDN|QHPOLGLUOHU6SHVLILNELUED÷ODQPDROPDGÕ÷Õ]DPDQ '1$NHVLPLJHQHOGHNoNGL]LOHUOH\DGDED]|]JQO÷\OHPH\GDQDJHOLU36 Bunun DNVLQHDONLOOH\LFLDMDQODUOD'1$NHVLPLELUYH\DGDKDID]OD'1$ED]ÕLoLQVHOHNWLYLWH gösterir. ùHNLO'1$úHNHULQGHQKLGURMHQDWRPXNRSDUÕOPDVÕ55¶ '1$RPXUJDVÕ O₂

.

O O O H B OR' R + RH RO O B OR' RO O B OR' RO RO O _B HO-O OR' O O B O RO -OR' O O RO B O Criegee Yeniden Düzenlenmesi +

(19)

'1$øOH(WNLOHúHQøODoODU

'1$ LOH HWNLOHúHQ LODoODUÕ o DQD EDúOÕN DOWÕQGD WRSOD\DELOLUL] '1$µGDNL ED] oLIWOHULDUDVÕQDJLUHUHN.-KHOLNV\DSÕ\ÕER]DQLQWHUNDODW|UOHU'1$ED]ODUÕ\ODNRYDOHQW RODUDNED÷ODQDQDONLOOH\LFLDMDQODUYHUDGLNDOROXúWXUDUDN'1$SROLQNOHRWLG]LQFLULQGH NHVLPHQHGHQRODQ'1$]LQFLULQLNÕUDQODU10

'1$øQWHUNDODW|UOHUL

Düz, genellikle aromatik ya da heteroaromatik moleküller .-KHOLNV\DSÕGDNLED] oLIWOHUL DUDVÕQD JLUHUHN '1$¶ \D ED÷ODQÕUODU YH ED] oLIWOHUL\OH NPHOHQLUOHU øQWHUNDODV\RQGDHWNLQRODQNXYYHWOHU\NWUDQVIHUNXYYHWOHULGLUIDNDWKLGURMHQED÷ODUÕYH HOHNWURVWDWLN NXYYHWOHU GH NDUDUOÕOÕNWD URO R\QDUODU37

øON RODUDN \ÕOÕQGD /HUPDQ WDUDIÕQGDQDoÕNODQDQLQWHUNDODV\RQLODFÕQKHOLNVLQHNVHQLQHULMLWELUúHNLOGHGLNRODUDN NRYDOHQWROPD\DQED÷ODQPDVÕGÕU38_{Bu da baz çiftlerini}_{QGúH\RODUDND\UÕOPDVÕQDVHEHS}

ROXU GROD\ÕVÕ\OD úHNHU IRVIDW RPXUJDVÕ ENOU YH KHOLNVLQ \DSÕVÕ ER]XOXU *|UQúH J|UH LQWHUNDODV\RQJoOH LOJLOLGLU %D]oLIOHULLOHLQWHUNDODW|UPROHNODUDVÕQGDNLYDQ GHU :DDOV NXYYHWOHUL ELUELUOHUL\OH NPHOHQPLú ED] oLIWOHUL DUDVÕQGDNL YDQ GHU :DDOV kuvvetleri’nden daha güçlüdür.39

ùHNLO Etidyum bromürün B-DNA’ya interkalasyonu. N+

N

H₂ NH₂

C₂H₅ Br

(20)

øQWHUNDODV\RQ :DWVRQ-&ULFN KLGURMHQ ED÷ODUÕQÕ NÕUPD] KHOLNV \DSÕ\Õ GHIRUPH HGHU'LUHN'1$KDVDUÕQDVHEHSROPD]KHOLNV\DSÕGDNRQIRUPDV\RQGH÷LúLNOL÷LQH\RO açar.

'1$LQWHUNDODW|UOHULQLoJUXEDD\ÕUDELOLUL]Akridinler (2), aktinomisinler (3) ve anthrasiklinler (4).10

(2) Amsacrine, R=R’=H (3) Actinomycin D, R=R’= D-Val

(4) Adriamycin, X=OH

1. 4. 2. Alkilleyici Ajanlar

'1$ DONLOOH\LFLOHUL LOH LQWHUNDODW|UOHUL DUDVÕQGDNL IDUN ED÷ODQPD úHNLOOHULGLU '1$LQWHUNDODW|UOHUL'1$LOHNRYDOHQWROPD\DQED÷ODUODED÷ODQÕUNHQDONLOOH\LFLDMDQODU NRYDOHQWED÷ODUODED÷ODQÕUODU.DQVHUNHPRWHUDSLVLQGHNXOODQÕODQHQ|QHPOLDONLOOHyici DMDQODU D]RW KDUGDOODUÕ HWLOHQLPLQOHU PHWDQRVOIRQLN DVLW HVWHUOHUL YH SODWLQ komplekleridir.10 N R R' HN NHSO2CH3 MeO L-Pro R' L-Thr O L-MeVal Sar L-Pro R' L-Thr O L-MeVal Sar O CH₃ CH₃ O N O O O O O OMe OH X O OH O H NH₃ OH C H₃

(21)

$]RW+DUGDOODUÕ

6OIUKDUGDOÕ,YH,,'Q\DVDYDúODUÕQGDNXOODQÕOPÕúoRNWRNVLNVLQLUJD]ÕGÕU, 'Q\D 6DYDúÕQGD VOIU KDUGDOÕ\OD |Oen askerlerin otopsileri sonucu lökopeni GúN EH\D] NDQ KFUHVL NHPLN LOL÷L aplazi, OHQI GRNXODUÕQGD ER]XQPD YH JDVWURLQWHVWLQDO VLVWHPGH OVHU VDSWDQPÕúWÕU %X OH]\RQODU VOIU KDUGDOODUÕQÕQ KÕ]OÕ E|OQHQ KFUHOHUH HWNLOL ROGX÷X VRQXFXQD J|WUPú YH DQWLWP|U HWNLOHUL RODELOHFH÷L GúQOPú \ÕOÕQGD VOIU KDUGDOÕ LQVDQODUGD WP|U KFUHVLQH HQMHNWH HGLOPLú40

IDNDW EX LúOHPLQ VLVWHPDWLNNXOODQÕPLoLQoRNWRNVLNROGX÷XDUWD\DoÕNPÕúWÕU*LOOPDQYHGL÷HUNLP\DFÕODU daha az toksik olan azot hardallaUÕQÕQDQWLWP|UHWNLVLQLLQFHOHPLúOHU\ÕOÕQGDD]RW KDUGDOÕQÕQ NOLQLN GHQHPHOHUL EDúODPÕúWÕU dDOÕúPD ,, 'Q\D VDYDúÕ VUHVLQFH GHYDP HWPLúWLU $]RW KDUGDOODUÕQÕQ \ÕOÕQGDQ EHUL NDQVHU WHGDYLVLQGH NXOODQÕOGÕ÷Õ bilinmektedir.41%XoDOÕúPDPRGHUQNDQVHUNHPRWHUDSLVLQLQEDúODQJÕFÕROPXúWXU

6OIUKDUGDOÕ $]RWKDUGDOÕ

5RVV¶D J|UH EL\RORMLN DONLOOH\LFL DMDQODU IL]\RORMLN NRúXOODUGD S+ 0 _C,

VXOXo|]HOWLKLGURMHQDWRPX\ODDONLOJUXEXQXQ\HULQLGH÷LúWLUHELOHQPROHNOOHUGLU'1$ için en iyiUHDNWLIRODQQNOHRILOLNNÕVÕPODU1-7 guanin > 3 adenin > 1 adenin > N-VLWR]LQVÕUDODPDVÕGÕU42

$]RW KDUGDOODUÕ ELIRQNVL\RQHO DONLOOH\LFL DMDQODUGÕU \DQL LNL WDQH HOHNWURILOLN NÕVÕPODUÕ YDUGÕU '1$ ED]ODUÕQÕ ]LQFLU LoL YH\D ]LQFLUOHU DUDVÕ ED÷OD\DEilirler. Azot KDUGDOODUÕQÕQ DONLODV\RQX úHNLO¶GDJ|VWHULOPLúWLU$]RWKDUGDOODUÕNRPúXLNL'1$ ]LQFLUL DUDVÕQD JLUHUHN LNL JXDQLQ ED]ÕQÕ 1-7 pozisyonundan alkilleyerek iki zinciri ELUELULQHED÷OD\DELOLUOHU10_ùHNLO

Cl S Cl Cl N Cl

(22)

ùHNLO Azot hardallaUÕ\ODDONLODV\RQ ùHNLO$]RWKDUGDOODUÕQÕQQHGHQROGX÷XoDSUD]ED÷ODQPD N R Cl N R Nu₁ Cl N R Nu₁ Cl Cl Nu₁ N R Nu₂ -Nu₂ N R Nu₁ + -+ + N N N N H O N H₂ N CH3 O O P O _O O O P O O O N N N NH O NH₂ O _O _P O O O O P O O O + -

(23)

-'L÷HU$ONLOOH\LFL$MDQODU

'L÷HUDONLOOH\LFLDMDQODU etileniminler (5), metanosülfonik asit esterleri (6), azot ND\QDNOÕODU7) ve platin kompleksleridir (8).10

(5) (6)

(7) (8)

'1$=LQFLULQL.ÕUDQODU

'1$ LOH HWNLOHúHQ LODoODUÕQ ED]ÕODUÕ EDúWD '1$ LQWHUNDODW|UOHUL JLEL ED]ODUÕQ DUDVÕQD JLUHUOHU GDKD VRQUD X\JXQ NRúXOODUGD UDGLNDOOHUL ROXúWXUXUODU %X UDGLNDOOHULQ '1$LOHHWNLOHúLPL'1$]LQFLULQLQNÕUÕOPDVÕQDVHEHSROXU

1. 4. 3. 1. Antrasiklin Antitümör Antibiyotikleri

'1$ LQWHNDODW|UOHUL RODQ DQWUDVLNOLQOHU D\QÕ ]DPDQGD RNVLMHQH ED÷OÕ '1$ KDVDUÕQDGDQHGHQROXUODU43 N N O R NHR' O H₃N Pt Cl Cl H₃N N N N N N N CH₃O₂SO-(CH₂) -OSO_n ₂CH₃

(24)

Doxorubicin, X = OH 1. 4. 3. 2. Bleomycin

%OHRP\FLQ\ÕOÕQGDStreptomyces verticillus’WDQL]ROHHGLOPLúELUDQWLWP|U DQWLEL\RWL÷LGLU44,45

Bleomycin

%OHRP\FLQGH SLULPLGLQ -DPLQRDODQLQ YH -hidroksiimidazol Fe(II) ile DNA NHVLP HWNLVL RODQ NDUDUOÕ ELU NRPSOHNV ROXúWXUXU 105 YH KLGURGLQDPLN oDOÕúPDODU \DSÕGDNL ELWKLD]ROXQ LQWHUNDODV\RQ HWNLVL ROGX÷XQX IDNDW EX LQWHUNDODV\RQXQ NODVLN LQWHUNDODW|UOHU HWLG\XP EURPU YE NDGDU HWNLVL ROPDGÕ÷ÕQÕ J|VWHUPLúWLU46,47_%D]Õ

oDOÕúPDODU ELWKLD]ROXQ WDP LQWHUNDOHWH ROPDGÕ÷ÕQÕ IDNDW '1$ \DSÕVÕQGDNL ROXNODUD ED÷ODQGÕ÷ÕQÕ J|VWHUPLúWLU48,49_{<DSÕGDNL VOIRQ\XP L\RQXQXQ IRVIDW JUXEX LOH}

O O O OMe OH X O OH O H NH₃ OH C H₃ N N H N NH₂ O N H₂ CH₃ O N H N H O H O O OH O H OH O O OH OH OH O O NH2 H NH2 O NH₂ H H H _H NH S N CH₃ H O N O NH S N O Me₂ S O H CH3 CH₃ H O H + H N NH

(25)

HOHNWURVWDWLN RODUDN HWNLOHúPH RODVÕOÕ÷Õ YDUGÕU50_{*OLNR] PDQQR] LVH EOHRPLFLQLQ ED]Õ}

kanser hücrelerinde selektif olarak birikiminden sorumlu olabilirler, fakat DNA kesiminde rol almazlar.

Bleomycin-Fe II Kompleksi

DNA kesim ürünlerinin incelenmesiyle, kesimin genellikle DNA zincirinin 3' XFXQGDNL SULQ QNOHRWLGLQH ED÷OÕ RODQ SULPLGLQ QNOHRWLQLQ NDUERQ DWRPXQGDQ KLGURMHQNRSDUÕOPDVÕ\ODJHUoHNOHúWL÷LDQODúÕOPÕúWÕU51

1. 4. 3. 3. Enediyen Antibiyotikleri

\ÕOÕQGD L]ROH HGLOHQ QHRFDU]LQRVWDWLQ52_{GÕúÕQGDNL HQHGL\HQ DQWLEL\RWLNOHUL}

esperamicin,53 calicheamisin,54 ve dynemicin A55 ¶OHULQ VRQODUÕQD GR÷UX oHúLWOL PLNURRUJDQL]PDODUGDQL]ROHHGLOPLúOHUGLU<DSÕODUÕQGDHQD]ELUoLIWED÷YHLNLoOED÷ EXOXQGXUGXNODUÕ LoLQ HQHGL\HQ DQWLWP|U DQWLEL\RWLNOHUL RODUDN DQÕOÕUODU %X WU moleküller DNA zincirinde küçük olukta interkalasyon yaparlar daha sonra SR veya 1$'3+JUXSODUÕ\ODDNWLYHHGLOHUHN'1$]LQFLULQLNHVHQUDGLNDOOHULQLROXúWXUXUOar.

1. 4. 3. 3. 1. Neocarzinostatin

\ÕOÕQGD Sterptomyces carzinostaticus’WDQL]ROHHGLOPLúWLU52_{Bilinen en eski}

HQHGL\HQDQWLEL\RWL÷LGLU=LQRVWDWLQRODUDNGDELOLQLU56_{7L\ROWDUDIÕQGDQDNWLYHHGLOHUHN}

Bergman, yeniden düzenlenmesiyle diradikal ROXúWXUXU 2NVLMHQ YDUOÕ÷ÕQGD LNL IDUNOÕ PHNDQL]PDLOH'1$NHVLPLJHUoHNOHúLU57-58 H NH S N CH₃ O N H O NH S N O Me₂S O H CH₃ CH₃ NH N N N H₂ O H NH2 N N H₂ O N H₂ CH₃ N H O H O-Seker O O H H + ' Fe N N H O₂

(26)

Neocarzinostatin

ùHNLO Neocarzinostatinin tiyol ile aktivasyonu

O C H₃ O H OH NHCH₃ CH₃ CH₃O OH O O O O O O O O C H₃ O H OH NHCH₃ O O O O O O Me OH O MeO SR -O C H₃ O H OH NHCH₃ Me OH O O MeO

. .

O O OH O O SR O C H₃ O H OH NHCH₃ Me OH O O MeO Bergman yeniden düzenlenmesi O O O O

.

(27)

ùHNLO$NWLYHHGLOPLúQHRFDU]LQRVWDWLQYHGL÷HUHQHGL\HQDQWLEL\RWLNOHULWDUDIÕQGDQ DNA keVLPPHNDQL]PDVÕ

1. 4. 3. 3. 2. Esperamicin ve Calicheamicin

(VSHUDPLFLQ YH FDOLFKHDPLVLQ DQWLWP|U DQWLEL\RWL÷LGLUOHU (VSHUDPLFLQ

Actinomadura verrucosospora, calicheamicin ise Micromonospora echinospora ssp. calichensis toprak örneklerinden izole ediOPLúOHUGLU54,60,61_{Neocarzinostatin gibi}

GLUDGLNDOROXúWXUDUDN'1$NHVLPLQHVHEHSROXUODU59 O B PO OP

.

O2 O B PO OP O-O

.

ROO

.

O B PO O OP O B OP O OH PO

..

+e, +H- + veya +RSH, -RS

.

PO H O PO H O DNA Kesimi O+ B OP O B OP OO

.

..

O B OP

.

+ H O₂ O OP H -O B Baz _OP H H O O + HO H O + PO -O

.

₂ -O2 DNA Kesimi

(28)

Esperamicin Calicheamicins O O O CH₃ O H O NH OMe MeO O C H₃ MeS OH O NHCH(CH₃)₂ OMe O O O C H₃ NH O NHCO₂CH₃ CH₃SSS O H O H O C H₂ MeO O C H₃ S OH O NHCH₂CH₃ OMe O O O C H₃ NH O NHCO2CH3 CH₃SSS O H O H O OH O H C H₃ MeO O CH3 I OMe OMe

(29)

1. 4. 3. 3. 3. Dynemicin A

 \ÕOÕQGD Micromonospora chersina’dan izole edilen dynemicin hem enediyen hem de antracyclinH DQWLEL\RWLNOHULQLQ \DSÕVDO RODUDN NRPELQDV\RQXGXU55

<DSÕVÕQGDNLDQWUDTXLQRQXQ'1$GXEOHNVLDUDVÕQDJLUHUHNLQWHUNDODV\RQDVHEHSROGX÷X WDKPLQ HGLOPHNWHGLU 'DKD VRQUD QHRFDU]LQRVWDWLQ JLEL GLUDGLNDO ROXúWXUDUDN '1$ úHNHULQGHQKLGURMHQDWRPXNRSDUDUDN'1A kesimine sebep olurlar.

Dynemicin A

1. 5. Karbon Merkezli Radikal Metabolitleri

Karbon merkezli radikaller normal metabolizmada yüksek konsantrasyonlarda ROXúPD]ODU61

Oksidatif stress62 ve karbonhidrat63 ya da amino asit64,65PHWDEROL]PDVÕQÕ etkiOH\HQ JHQHWLN G]HQVL]OLNOHU JLEL SDWRORMLN GXUXPODUGD ROXúXUODU 6RQXoWD NDUERQ merkezli radikaller karbontetraklorür,66,67 hidrazin türevleri,68 nitroalkanlar,69,70 etanol71,72, diazoquinonlar73,74 ve organik hidroperoksitler75,76JLELoHúLWOLNLP\DVDOODUÕQ PHWRELOL]PDVÕVRQXFXED]ÕJHQRWRNVLN|]HOOLNOHUHED÷OÕRODUDNROXúXUODU7DEOR 3ROLVLNOLN DURPDWLN KLGURNDUERQODUÕQ DURPDWLN NDW\RQ UDGLNDOOHULQH G|QúPHVL VHUEHVW radikal ara ürünlerine güzel bir örnek olarak verilebilinir.

Karbon merkezli radLNDOOHU PROHNOHU RNVLMHQ LOH oRN KÕ]OÕ HWNLOHúHUHN DONLO UDGLNDOLQGHQGDKDUHDNWLIELRPROHNOOHURODQSHURNVLWUDGLNDOLJLELUQOHUROXúWXUXUODU77

.LP\DVDO NDQVHURMHQOHUOH LOJLOL ELUoRN oDOÕúPD '1$¶\D NRYDOHQW ED÷ODQPDQÕQ kanserojenlerin alkildiazonyu_{P L\RQODUÕ \D GD HSRNVLWOHU JLEL HOHNWURILOLN DUD UQOHUL} DNWLYH HGHUHN L\RQLN PHNDQL]PD LOH ROGX÷X \|QQGHGLU78,79₍_{ùHNLO ) Karbon}

PHUNH]OLUDGLNDOOHULQ'1$LOHHWNLOHúHQUHDNWLIWUOHUROGX÷XELOLQLU80 N H O OH O CH3 CH₃ O O O OH OH

(30)

ùHNLOAlkil radikali ile DNA hasarÕ-dimetil hidrazin ile DNA baz ürünlerinin ROXúXPX CH₃-N-N-CH₃ Metabolizma C H₃ N N + _CH 3

.

Gua-DNA N N N N H O N H₂ R CH₃ N N N N H O N H₂ R CH₃ H N N N N H O N H2 R N-MeGua-DNA7 . + N N N N N H₂ R OMe N N N N H O N H₂ R CH₃ O-MeGua-DNA6 C-MeGua-DNA8 H H

(31)

Xenobiotik Serbest Radikal Metabolit Genotoksik Özellik CCl4 Hidrazinler Triklorometil Alkil, Aril ÜCCl3 RÜArÜ Hepatokanserojen81 Mutajen,kanserojen Etanol .-hidroksietil H3C- ÜCHOH Teratojen

2-Nitropropan .-hidroksiizopropil (H3C)2ÜCOH Mutajen

Hepatokanserojen81 Diazoquinonlar Peroksitler .-hidroksifenil alkil, peroksil, alkoksil ÜArOH RÜROOÜ ROÜ Mutajen74,75 Kanserojen82,83 Promoter75,76

PAHa Aril katyonu Ar+-ÜCH Kanserojen84

Tablo 1. 1. *HQHWRNVLN ;HQRELRWLNOHUGHQ ROXúDQ NDUERQ PHUNH]OL VHUEHVW UDGLNDO metabolitleri.

a_{Polisiklik aromatik hidrokarbonlar}

Alkil radikalleri hidroksil radikalinden daha az reaktiftirler. Hem alkil radikali KHP GH KLGURNVLO UDGLNDOL '1$ úHNHULQGHQ KLGUojen atomu kopararak ya da DNA ED]ODUÕ\ODHWNLOHúHUHN]LQFLUNHVLPLQH\DGDDONLODV\RQDVHEHSROXUODU85

ùHNLO

ùHNLO.DUERQPHUNH]OLUDGLNDOPHWDEROLWOHULLOHROXúDQ'1$PRGLILNDV\RQúHPDVÕ DNA + R

.

Pürin baz modifikasyonu

Fosfodiester bag kesimi N N N N H O N H₂ R N N N HN NH₂ R ( 8 - alkil pürin)

(32)

g1&(.ød$/,ù0$/$5

/LWHUDWU WDUDPDVÕ o \|QGH JHUoHNOHúWLULOGL $ULO UDGLNDOOHUL YH '1$ NHVLPL '1$NHVLPLQLQ|QOHQPHVLYHELWNLHNVWUDNWODUÕLOH'1$NHVLPL

gQFHNLoDOÕúPDODUGDDULOUDGLNDOOHULQLQ'1$NHVLPLQH\RODoWÕ÷ÕYHGLD]RQ\XP WX]ODUÕQÕQ ELU HOHNWURQ GRQ|U YDUOÕ÷ÕQGD '1$¶\Õ NHVWL÷L UDSRU HGLOPLúWLU86_$\QÕ

]DPDQGD ED]Õ ELWNL HNVWUDNODUÕQÕQ YH\D ELWNLOHUGHQ L]ROH HGLOHQ ELOHúLNOHULQ '1$ NHVLPLQL |QOHGL÷L \LQH ED]Õ ELWNL HNVWUDNWODUÕQÕQ YH\D ELWNLOHUGHQ L]ROH HGLOHQ ELOHúLNOHULQ &X+2_{YDUOÕ÷ÕQGD '1$ NHVLPLQH \RO DoWÕ÷Õ GD \DSÕODQ OLWHUDWU oDOÕúPDVÕ}

VRQXFX WHVSLW HGLOPLúWLU87_{g]HOOLNOH \HQLOHELOHQ ELU NOWU PDQWDUÕ RODQ Agaricus}

bisporus¶XQ SDUoDODQPD UQOHULQLQ IDUHOHUGH NDQVHURMHQ HWNL\H VDKLS ROPDVÕ YH EX

SDUoDODQPD UQOHULQGHQ ELULQLQ EHQ]HQGLD]RQ\XP L\RQX ROPDVÕ88 _{EL]L GH÷LúLN}

diazonyumWX]ODUÕVHQWH]OH\LSEXQODUÕQ'1$NHVLPHWNLVLQLLQFHOHPH\H\|QHOWPLúWLU $\UÕFD DQWLRNVLGDQW |]HOOL÷L ELOLQHQ ELWNL HNVWUDNODUÕQÕQ ELU NÕVPÕQÕQ '1$ kesimini önlemesi antioksidant ve antimikrobiyal özellikleri belirlenen89 ED]Õ

Hypericum ve Achillea türü bitkilerin DNA kesimini önleme kapasitesine bakmaya

\|QHOWPLúWLU

Aril Radikali ve DNA Kesimi

*ULIILWKV YH $UN \DSPÕú ROGXNODUÕ oDOÕúPDGD ELU HOHNWURQ GRQ|U YDUOÕ÷ÕQGD EHQ]HQGLD]RQ\XP WHWUDIORURERUDWÕQ '1$¶\Õ NHVWL÷LQL J|VWHUPLúOHUGLU %X oDOÕúPDGD bHQ]HQGLD]RQ\XPVXGDL\Lo|]QPHVLYHSR]LWLI\NOROPDVÕQGDQGROD\Õ'1$NHVLPL LoLQJ]HOELUPRGHORODELOHFH÷LGúQOPú'LD]RQ\XPWX]ODUÕSR]LWLI\NOROGXNODUÕ LoLQ '1$ \DSÕVÕQGDNL QHJDWLI \NO IRVIDW JUXSODUÕ\OD HWNLOHúLUOHU (OHNWURQ GRQ|U YDUOÕ÷ÕQGDD]RWXQD\UÕOPDVÕ\ODDULOUDGLNDOLROXúXU$ULOUDGLNDOLGHGHRNVLULER]úHNHUGHQ KLGURMHQDWRPXNRSDUDUDN'1$NHVLPLQLJHUoHNOHúWLULU86 ùHNLO$ULOUDGLNDOLROXúXPPHNDQL]PDVÕ N N N N N N + +

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e

(33)

%DúND ELU oDOÕúPDGD \HQLOHELOHQ PDQWDUGD EXOXQDQ -(hidroksimetil)benzendiazonyum tuzunun (9 ROXúWXUGX÷X NDUERQ PHUNH]OL UDGLNDOLQ '1$ ED]ODUÕ YH GHRNVLULER] úHNHUL\OH PRGLILNDV\RQX oDOÕúÕOPÕúWÕU -KLGURNVLPHWLOEHQ]HQGLD]RQ\XP WX]XQXQ '1$ \DSÕVÕQGD EXOXQDQ ED]ODU LOH 1-8 SR]LV\RQXQGDQ ED÷ODQDUDN -HMP-dGuo (10) ve 8-HMP-dAdo (11 ROXúWXUGX÷XQX GHRNVLULER]úHNHULOHGHPDORQGLDOGHKLWROXúWXUGX÷XQXUDSRUHGLOPLúWLU88

(9) (10) (11)

5HV]ND YH DUN GLD]RQ\XP WX]ODUÕ 12a-d)’yi kullanarak, diazonyum ELOHúLNOHULQLQHOHNWURQGRQ|UYDUOÕ÷ÕQGDDULOUDGLNDOLQHG|QúW÷Q(35VSHNWUXPODUÕQÕ LQFHOH\HUHN EXOPXúODUGÕU %X oDOÕúPD VXOX o|]HOWLGH DULO UDGLNDOL HOGH HWPHN LoLQ DUHQGLD]RQ\XPL\RQODUÕQLQGLUJHQPHVLQLQX\JXQPHWRGROGX÷XQXJ|VWHUPLúWLU6RQXoWD EX DUDúWÕUPD ELRORMLN LQdirgeyici maddelerin ArN2+’nu parçalayarak aril radikaline

G|QúWUPH\HWHQH÷LQHVDKLSROGX÷XQXGR÷UXODU90 (12) a) X =MeO b) X = Cl c) X = Br d) X = NO2 e) X = N(Et)2 N N OH + O H N N N N H O N H₂ CH₂OH O _H_O N N N N NH₂ CH₂OH O X N+₂ BF-₄

(34)

%DúNDELUoDOÕúPDGD'HY3$U\DYHDUNIDUNOÕLNLDULOKDONDVÕQGa diazonyum grubu içeren 1,4-ELVGLD]RQ\XP ELOHúLNOHULQLQ EDNÕU NORUU YDUOÕ÷ÕQGD -GLUDGLNDOOHUL ROXúWXUDUDN HQHGL\HQ DQWLEL\RWLNOHUL JLEL '1$¶\Õ NHVWL÷LQL J|VWHUPLúOHUGLU91

(13)

Yine Dev P. Arya ve ark., 9-'LD]RIOXRUHQYH-naftal fenil diazometan (15) ELOHúLNOHULQLQ EDNÕU DVHWDW YDUOÕ÷ÕQGD RNVLMHQOL RUWDPGD S%5 SOD]PLG '1$¶\Õ NHVWL÷LQLJ|VWHUPLúOHUGLU5HDNVL\RQODUÕNDUDQOÕNWD\DSWÕNODUÕLoLQNHVLPPHNDQL]PDVÕQÕQ \D NDUERQ PHUNH]OL UDGLNDOLQ '1$ GHRNVLULER] úHNHULQGHQGHQ KLGURMHQ DWRPu NRSDUPDVÕ\OD \D GD EDNÕU DVHWDWÕQ '1$ NHVLP HWNLVL RODQ &X,-Oksijen kompleksi YH\D KLGURNVLO UDGLNDOL JLEL DNWLI RNVLMHQ WUOHUL ROXúWXUDUDN JHUoHNOHúWL÷LQL UDSRU HWPLúOHUGLU92 (14) (15) %DúNDELUoDOÕúPDGDJHQHO\DSÕODUÕp-X-Ar+ _{RODQGLD]RQ\XPWX]ODUÕQGDQS-X-Ar}. ROXúDELOHFHNNRúXOODUGDFW'1$\DGDKUHOHUOHHWNLOHúLPLVRQXFXJXDQLQ'1$UQOHUL ROXúWX÷XUDSRUHGLOPLúWLUp-X-Ar

Ü

ROXúXPX(65LOHDoÕNODQPÕúWÕU93 ùHNLO$UHQGLD]RQ\XPL\RQODUÕYH'1$¶GDQHOGHHGLOHQ&-arilguanin ürünleri (X = CH3, CH2OCH3, CH2OH) H N H N O N H O O N N₂ N₂ H O N H N H + + + + N+₂ _ N+₂ X N+₂ HSO₄ HN N N N X H N H₂

(35)

:DUQHU YH DUN VEVWLWH WULDULODPLGOHU VHQWH]OH\HUHN EXQODUÕQ '1$ NHVLP HWNLVLQLQLQFHOHPLúOHU.HVLPHWNLVLQLQD!F!EROGX÷XQXJ|]OHPOHPLúOHU94 (16) a) Y = H b) Y = NO c) Y = OMe ùHNLO6EVWLWHWULDULODPLGOHULQ'1$¶\ÕNHVLPL

%DúND ELU oDOÕúPDGD %XUU YH DUN ELU %HUHQLO DQDOR÷X RODQ N-(3-Hydroxypropyl)-Berenil (18 ELOHúL÷L YH -amidinobenzendiazonyum tuzunun (17) '1$ NHVLPLQL LQFHOHPLúOHU YH EX ELOHúLNOHULQ '1$¶\Õ NHVWL÷LQL J|]OHPOHPLúOHU 18) ELOHúL÷LQLQ SDUoDODQDUDN 17 ELOHúL÷LQL ROXúWXUGX÷XQX YH E|\OHFH NHVLPLQ EDúODGÕ÷ÕQÕ UDSRUHWPLúOHUGLU95

ùHNLO N-(3-Hydroxypropyl)-Berenil sentezi (17) (18) CONH O₂N NHCO CO₂Et Y NH₂ NH₂ CN O H Br N CN H NH₃ N NH H₃N OH H OH N H H3N N₂ NH N H N N N NH₃ NH₃ OH + + + ₊ MeOH/HCl /MeOH %50 + %65 %50

(36)

.Õ]ÕO YH DUN EHQ]HQGLD]RQ\XP WHWUDIORURERUDW 19a-c WX]ODUÕQÕ Ventezleyip '1$NHVLPLQLLQFHOHPLúOHU6RQXoRODUDNIDUNOÕGLD]RQ\XPWX]ODUÕQÕQELUHOHNWURQGRQ|U YDUOÕ÷ÕQGD '1$¶\Õ NHVWL÷LQL J|]OHPOHPLúOHU .HVLP HWNLVL LQFHOHQHQ GLD]RQ\XP WX]ODUÕQÕQ UDGLNDO DNWLYLWHOHUL YH '1$ NHVLP HWNLOHUL DUDVÕQGD DQODPOÕ ELU IDUNOÕOÕN J|]OHQPHGL÷LQLUDSRUHWPLúOHUGLU96

19 a) R = H b) R = OH c) R = CO2H

DNA Kesiminin Önlenmesi

$WWDJXLOHYHDUNKDONDUDVÕQGDoHúLWOLGHULKDVWDOÕNODUÕQÕQWHGDYLVLQGHD\UÕFDD÷UÕ kesici olarak ve antidiaretik ilaç olarak kulODQÕODQ Cistus incanus L. ve Cistus

monspeliensis /¶QLQ ELWNL HNVWUDNODUÕQÕQ DQWLRNVLGDQW YH '1$ NHVLP HWNLOHULQL

DUDúWÕUPÕúODUGÕU %X WU ELWNLOHU IODYRQRLG LoHULUOHU YH '1$ ]LQFLULQL NÕUDELOHFH÷L GúQOHQDQWLRNVLGDQWPDGGHOHUGLU%XELWNLHNVWUDNWODUÕQÕQ'1$NHVLPLQL|QOHGL÷LYH GR]DED÷OÕRODUDNUDGLNDOV|QGUPHNDSDVLWHVLROGX÷XQXJ|]OHPOHPLúOHUGLU97

<LQH EDúND ELU oDOÕúPDGD ELRIODYRQRLGOHULQ DQWLUDGLNDO DQWLRNVLGDQW YH '1$ NHVLPLQL|QOHPHNDSDVLWHVLLQFHOHQPLúWLU'R÷DOSROLIHQROLNELOHúLNOHURODn flavonoidler LQVDQODU WDUDIÕQGDQ WNHWLOHQ PH\YHOHUGH YH VHE]HOHUGH EXOXQXUODU (Q E\N ND\QDN HOPDVR÷DQGXWJLEL\L\HFHNOHUYHoD\JLELLoHFHNOHUGLU6RQXoRODUDNELRIODYRQRLGOHULQ UXWLQFDWHFKLQYHQDULQJLQGR]DED÷OÕRODUDNVHUEHVWUDGLNDOOHULsöndürdükleri ve DNA NHVLPLQL|QOHGLNOHULUDSRUHGLOPLúWLU98

<LQH5XVVRYHDUN+LQWúLIDOÕELWNLOHULQLQDQWLUDGLNDOYH'1$NHVLPLQL|QOHPH NDSDVLWHOHULQL DUDúWÕUPÕúODUGÕU Celastrus paniculatus L., Picrorhiza kurroa L. ve

Withania somnifera L., epilepsLX\NXVX]OXN UDúLWL]PJXWYHKDVÕPVÕ]OÕNWHGDYLVLQGH NXOODQÕODQ úLIDOÕ ELWNLOHUGLU %X oDOÕúPDGD ELWNLOHULQ PHWDQRO HNVWUDNODUÕQÕQ +2O2’tin

BF₄N₂

R

(37)

UV-IRWROL]LVRQXFXROXúDQ'1$KDVDUÕ]HULQGHNLHWNLVLLQFHOHQPLúWLU%WQHNVWUDNODU GR]D ED÷OÕ RODUDN VHUEHVt radikal söndürme ve DNA kesimini önleme etkisi J|VWHUPLúOHUGLUPicrorhiza kurroa/¶QLQPHWDQROHNVWUDNWÕQÕQGL÷HUELWNLHNVWUDNWODUÕQD J|UHGDKDHWNLOLROGX÷XJ|]OHQPLúWLU99

%DúND ELU oDOÕúPDGD NÕUPÕ]Õ SRUWDNDO HNVWUDNWÕQÕQ '1$ NHVLPL ]HULQH HWNLVL LQFHOHQPLú.ÕUPÕ]ÕSRUWDNDOODUIODYRQODUKLGUR[\FLQQDPLFDVLWOHUYHDVNRUELNDVLWJLEL DQWLRNVLGDQW ELOHúLNOHULQ \DQÕQGD \NVHN RUDQGD DQWURF\DQLQOHU GH LoHULUOHU (NVWUDNW '1$ NHVLPL ]HULQGH NRUX\XFX HWNL YH GR]D ED÷OÕ RODUDN GD UDGLNDO V|QGUPH kapasLWHVLJ|VWHUPLúWLU100

$FTXDYLYDYHDUN5HVYHUDWURO¶Q'1$NHVLPL]HULQGHNLHWNLVLQLLQFHOHPLúOHU 5HVYHUDWURO úDUDSWD EXOXQDQ GR÷DO SK\WRDOH[LQ PDGGHVLGLU YH GDPDU KDVWDOÕNODUÕQGD NRUX\XFXHWNLVLROGX÷XELOLQLU%XoDOÕúPDGD5HVYHUDWURO¶QGR]DED÷OÕolarak serbest UDGLNDO V|QGUPH NDSDVLWHVL YH '1$ NHVLPLQL NRUX\XFX HWNL J|VWHUGL÷L WHVSLW HGLOPLúWLU101

:DQJYHDUNDGDúODUÕ/RWXV3OXPXOHYH%ORVVRPNelumbo nucifera Gertn.)’um DQWLRNVLGDQWHWNLVLLQFHOHQPLú/RWXVSOXPXOHoHúLWOLDONDORLGOHULoHULUYHKDONDUDVÕQGD DQWLKLSHUWDQVLI LODo RODUDN NXOODQÕOÕU /RWXV EORVVRP LVH DONRORLGOHU RUJDQLN DVLWOHU DPLQR DVLWOHU YH -carotenoidler içerir. Hidroksil radikali söndürme etkisi LQFHOHQGL÷LQGH /RWXV 3OXPXOH YH %ORVVRP¶XQ GR]D ED÷OÕ RODUDN KLGURNVLO UDGLNDlini V|QGUG÷UDSRUHGLOPLúWLU102

Yine Acquaviva ve ark., cyanidin ve cyanidin 3---D-glikozid’in DNA NHVLPLQL |QOHPH NDSDVLWHVL YH DQWLRNVLGDQW |]HOOL÷LQL LQFHOHPLúOHUGLU dDOÕúPDGD cyanidin ve cyanidin 3---D-glikozid’in DNA kesimi üzerine koruyucXHWNLVLROGX÷X YHGR]DED÷OÕVHUEHVWUDGLNDOOHULV|QGUG÷J|]OHQPLúWLU103

(38)

(20) R = H, Cyanidin

R = Glikoz, 3---D-glikozid

Russo ve ark., Bacopa monniera/¶QÕQVHUEHVWUDGLNDOV|QGUPHNDSDVLWHVLQLYH '1$KDVDUÕQÕ|QOHPHHWNLVLQLDUDúWÕUPÕúODUGÕUBacopa monniera/NOLQLNRODUDNKDIÕ]D ND\EÕQGD HSLOHSVLGH X\NXVX]OXN SUREOHPLQGH YH D÷UÕ NHVLFL RODUDN NXOODQÕOÕU dDOÕúPDGDBacopa monniera/¶QLQPHWDQROHNVWUDNWÕNXOODQÕOPÕúWÕU'R]DED÷OÕRODUDN KLGURNVLOUDGLNDOLQLV|QGUG÷YH'1$NHVLPLQL|QOHGL÷LUDSRUHGLOPLúWLU104

Tian ve ark., aloin (21) ve aloe-emodin’in (22 NRQVDQWUDV\RQD ED÷OÕ RODUDN SURRNVLGDQW YH DQWLRNVLGDQW HWNLOHULQL LQFHOHPLúOHUGLU <NVHN NRQVDQWUDV\RQODUGD DORLQ¶LQKLGURNVLOUDGLNDOLQHED÷OÕRODQ'1$NHVimini %5-RUDQÕQGD|QOHGL÷LWDP WHUVLQH GúN NRQVDQWUDV\RQODUGD LVH NHVLPL DUWWÕUGÕ÷Õ J|]OHQPLúWLU $ORH-emodin’in GúN NRQVDQWUDV\RQODUGD oRN D] DQWLRNVLGDQW HWNL J|VWHUGL÷L \NVHN konsantrasyonlarda ise prooksidant etki göstererek DNA kesimini %29- RUDQÕQGD D]DOWWÕ÷ÕWHVSLWHGLOPLúWLU105 (21) (22) O OH OH OR OH O H + OH O OH O CH₂OH O OH O OH CH₂OH OH OH O H CH2OH

(39)

%DúND ELU oDOÕúPDGD 5RRLERV oD\ÕQÕQ Aspalathus linearis) antioksidant DNWLYLWHVL YH '1$ NHVLPLQL |QOHPH NDSDVLWHVL DUDúWÕUÕOPÕúWÕU dDOÕúPDGD HWDQRO YH VX HNVWUDNWODUÕ NXOODQÕOPÕúWÕU +HP HWDQRO HNVWUDNWÕQÕQ KHP GH VX HNVWUDNWÕQÕQ SR]LWLI NRQWURORODQ7UROR[LOHKHPHQKHPHQD\QÕRUDQGDKLGURNVLOUDGLNDOLQGHQND\QDNODQDQ '1$NHVLPLQL|QOHGL÷LJ|]OHQPLúWLU106

<HQYHDUNORWXVVHHGHNVWUDNWÕQÕQDQWLRNVLGDQW|]HOOLNOHULQLYHLQVDQOHQIRVLWOHUL üzerLQGHNL '1$ NHVLP HWNLVLQL DUDúWÕUPÕúODUGÕU /RWXV VHHG¶LQ ND\QDWÕOPÕú VX HNVWUDNWÕQÕQ JoO DQWLRNVLGDQW HWNL J|VWHUGL÷L EXOXQPXúWXU 6X HNVWUDNWÕQÕQ OLSLW SHURNVLGDV\RQX YH '1$ KDVDUÕ ]HULQGH KHP KLGURMHQ SHURNVLWOL RUWDPGD KHP GH hidrojen peroksitsiz orWDPGDKLoELUHWNLVLROPDGÕ÷ÕJ|]OHQPLúWLU107

<DPDJXFKL YH DUN %UH]LO\D oDPÕ Araucaria angustifolia¶QÕQ LoHUGL÷L ELRIODYRQRLGOHULQ '1$ KDVDUÕ ]HULQGHNL NRUXPD HWNLVLQLQ LQFHOHPLúOHU Araucaria

angustifolia’da bulunan 6 tane biflavonoidi HPLC/MS ile aQDOL]HWPLúOHU'R]DED÷OÕ olarak Araucaria angustifolia¶QÕQ'1$NHVLPLQL|QOHGL÷LQLEXOPXúODUGÕU108

R1 = R2 = R3 = R4 = OH amentoflavone Mono-O-methylamentoflavone Di-O-methylamentoflavone R1 = R2 = R3 = R4 = OMe ginkgetin Tri-O-methylamentoflavone R1 = R2 = R3 = R4 = OMe tetra-O-methylamentoflavone ùHNLO%UH]LO\DoDPÕAraucaria angustifolia¶QÕQLoHUGL÷LELIODYRQRLGOHU O R OH O O R4 OH O R 1 3 2 R

(40)

$KPDG YH DUN ELWNL ND\QDNOÕ ELU SROLIHQROLN ELOHúLN RODQ UHVYHUDWUROQ SURRNVLGDQWDNWLYLWHVLQLEDNÕUL\RQXYDUOÕ÷ÕQGDLQFHOHPLúOHU5HVYDUDWUROQDQWLNDQVHUYH NDUGL\RYDVNOHU|]HOOLNOHULQLQROPDVÕELUoRNDUDúWÕUPDFÕODUÕQEXPROHNOHRODQLOJLVLQL DUWWÕUPÕúWÕU %X oDOÕúPDGD UHVYDUDWUROQ EDNÕU L\RQX YDUOÕ÷ÕQGD SURRNVLGDQW |]HOOL÷L ROGX÷XJ|VWHULOPLúWLU5HVYHUDWUROQEDNÕUL\RQXYDUOÕ÷ÕQGDSOD]PLG'1$¶GDPXWDV\RQD VHEHSROGX÷XWHNEDúÕQDLVHELUoRN*XDQLQED]ÕQÕVLOGL÷LEHOLUOHQPLúWLU87

%LWNLOHUøOH'1$.HVLPL

%LUGR÷DOUQRODQ5HVYHUDWURO¶Q-trihidroksi-trans-stilben, 23EDNÕU,, NORUU YH RNVLMHQ YDUOÕ÷ÕQGD S%5 SOD]PLG '1$¶\Õ NHVWL÷L J|VWHULOPLúWLU 'L÷HU PHWDOOHU YDUOÕ÷ÕQGD 5HVYHUDWURO¶Q '1$ NHVLPLQL EDúODWPDGÕ÷Õ için Cu+2_{‘nin DNA}

NHVLPLQGH5HVYHUDWUROVSHVLILNPHWDOL\RQXROGX÷XUDSRUHGLOPLúWLU86

(23)

Hadi ve ark., serotoninin (24) Cu+2 varlÕ÷ÕQGD'1$NHVLPHWNLVLQLLQFHOHPLúOHU Serotonin (5-hydroxytryptamine) beyinde ve omurilikte önemli bir nörotransmitterdir; X\NX ELOLQo KX\VX]OXN YH VDOGÕUÕ NRQWUROQGH J|UHY DOÕU %X oDOÕúPDGD 6HURWLQLQ¶LQ Cu+2 YDUOÕ÷ÕQGD RNVLGDWLI PHNDQL]PD LOH '1$ NHVLPLQH VHEHS ROGX÷X EHOLUOHQPLúWLU 6HURWRQLQ&X,,¶\L&X,¶LQGLUJHUYHEDNÕUL\RQXYDUOÕ÷ÕQGDKLGURNVLOUDGLNDOLROXúXU110 (24) OH OH O H Cu , O2+ ₂ pH ,7 2 DNA Kesimi N H O H NH₂

(41)

8GGLQYHDUNELIODYRQRLGRODQDPHQWRIODYRQHQXQEDNÕUL\RQXYDUOÕ÷ÕQGD'1$ NHVLPLQHVHEHSROGX÷XQXUDSRUHWPLúOHUGLU. Ametoflavone Cu(II)’yi Cu(I)’ indirger ve EDNÕU L\RQX YDUOÕ÷ÕQGD KLGURNVLO UDGLNDOL ROXúXU $\UÕFD VSHNWURVNRSLN YHULOHU &X,, YDUOÕ÷ÕQGD DPHWRIODYRQHQXQ 25 EDNÕU L\RQXQD ED÷ODQPD NDSDVLWHVL ROGX÷XQX J|VWHUPLúWLU111

(25)

Ma ve ark. Alangium javanicum’dan izole ettikleri Javaniside’in (26) Cu2+ YDUOÕ÷ÕQGD'1$NHVLPLQLEDúODWWÕ÷ÕQÕWHVSLWHWPLúOHUGLU112

(26)

Ma ve ark., Piper caninum¶XQKDPHNVWUDNWÕQÕQS%5SOD]PLG'1$¶\Õ&X+2

YDUOÕ÷ÕQGD NHVWL÷L EHOLUOUQPLúWLU 'DKD VRQUD HNVWUDNW IUDQNVL\RQODQGÕUÕODUDN LQ YLWUR

RODUDN'1$NHVLPHWNLOHULLQFHOHQPLúYHoWDQHDNWLIELOHúLNL]ROHHGLOPLú%XQODUN-cis-feruloyl tyramine (27), N-trans-feruloyl tyramine (28) ve 1-cinnamoylpyrrolidine’dir (29). 27-29ELOHúLNOHUL'1$NHVLPLJHUoHNOHúWLUHQPDGGHOHr LoHULVLQGH\DSÕVDORODUDN\HQLELUVÕQÕIÕWHPVLOHGHUOHU113 O O H OH O M O 2+ N H O N O O H _H O O OH OH CH₂OH O H

(42)

(27) (28)

(29)

0DYHDUNEDúNDELUoDOÕúPDODUÕQGD\LQHPiper caninum’un DNA kesim etkisi gösteren bir fraksiyonundan 4,5-dioxoaporphine alkoloid cepharadione A (30ELOHúL÷LQL izROHHWPLúOHUGLU114

(30)

Yine Ma ve ark., Malotus resinosus¶XQN|NOHULQGHQKD]ÕUODGÕNODUÕKDPHNVWUDNWÕQ Cu+2YDUOÕ÷ÕQGD'1$NHVLPHWNLVLROGX÷XQXEHOLUOHPLúOHU'DKDVRQUD'1$NHVLPLQLQ basit bir coumarin olan scopoletin (31a ELOHúL÷LQGHQ ND\QDNODQGÕ÷ÕQÕ UDSRU HWPLúOHU S%5SOD]PLG'1$NXOODQDUDNNHVLPHWNLVLQLLQYLWURRODUDNJ|]OHPOHPLúOHU31a) ELOHúL÷LQLQ GDKD DNWLI RODFD÷ÕQÕ GúQGNOHUL o DQDOR÷XQXQ GD '1$ NHVLP HWNLVLQL LQFHOHPLúOHU'1$NHVLPHWNLOHULQLQ31c>31a=31b>31dROGX÷XQXUDSRUHWPLúOHUGLU115 N H O OH O H MeO O H MeO N H O OH N O O O N O O

(43)

(31) a) R1 = CH3 R2 = H b) R1 = H R2 = CH3 c) R1 = H R2 = H d) R1 = CH3 R2 = CH3 O O R1O R₂O

(44)

3. MATERYAL VE METOT 3. 1. Materyal

%X oDOÕúPDQÕQ PDWHU\DOLQL ROXúWXUDQ ELWNL |UQHNOeri Dicle Üniversitesi, Fen (GHEL\DW )DNOWHVL %L\RORML %|OP¶QGHQ 3URI 'U $ 6HOoXN (57(.ø1 WDUDIÕQGDQ WRSODQGÕYHWHúKLVHGLOGL7HúKLVHGLOHQELWNL|UQHNOHUL'LFOHhQLYHUVLWHVL)HQ(GHEL\DW )DNOWHVL+HUEDU\XPXQGD'8)VDNODQPDNWDGÕU

3. 1. 1. KXOODQÕODQ%LWNL7UOHUL7DQÕPODUÕ<D\ÕOÕúODUÕYH.XOODQÕP$ODQODUÕ

Hypericum retusum, 'L\DUEDNÕU dQJú dHUPLN-dQJú NP WDUOD

NHQDUÕQGDQWDULKLQGHWRSODQGÕ1R'8)

Hypericum scabrum, 'L\DUEDNÕU (UJDQL (UJDQL 0DGHQ NP \DPDoODUdan

WDULKLQGHWRSODQGÕ1R'8)

Hypericum lysimachioides, 'L\DUEDNÕU (UJDQL (UJDQL-Çermik 11 km,

\DPDoODUGDQWDULKLQGHWRSODQGÕ1R'8)

Achillea aleppica subsp. aleppica,'L\DUEDNÕU'L\DUEDNÕU-Silvan 38 km, yol NHQDUÕQGDQWDULKLQGHWRSODQGÕ1R'8)

Achillea aleppica subsp. zederbaueri,

0DUGLQ0D]ÕGD÷Õ<XNDUÕ.RQDNN|\-NP\DPDoODUGDQWDULKLQGHWRSODQGÕ1R'8)

Achillea biebersteinii 'L\DUEDNÕU 'L\DUEDNÕU-6LYHUHN NP \RO NHQDUÕndan

WDULKLQGHWRSODQGÕ1R'8)

Hypericum WUOHUL oRN \ÕOOÕN RWVX ELWNLOHUGLU hONHPL]GH WU EXOXQDQ Hypericum¶XQ WU *QH\GR÷X $QDGROX %|OJHVLQGH \D\ÕOÕú J|VWHUPHNWHGLUOHU

.DQWDURQ NDQWDUXP NR\XQNÕUDQ YH ELQELUGHOLN RWX RODUDN bilinirler. Geleneksel WHGDYLGH \DSUDNOÕ oLoHNOL YH PH\YHOL GDOODUÕ LOH N|NOHUL NXOODQÕOÕU 0LGH UDKDWVÕ]OÕNODUÕQGDLQIODPDV\RQGDYHGHSUHV\RQWHGDYLVLQGHNXOODQÕOÕUODU116_Hypericum

WUOHULIODYRQRLGOHUYHWHPHO\D÷DVLWOHULLoHULUOHU

Achillea türleri oRN\ÕOOÕNW\VYHRWVXELWNLOHUGLUhONHPL]GHWUEXOXQXUYH

EXQODUGDQ WU *QH\GR÷X $QDGROX %|OJHVLQGH \D\ÕOÕú J|VWHUPHNWHGLUOHU &LYDQSHUoHPLDNEDúOÕEDUVDPRWXELQELU\DSUDNRWXPDUVDPRWXNDQGLOoLoH÷LRODUDN bilinirler. Geleneksel tedaviGH UDKLP KDVWDOÕNODUÕQGD LGUDU V|NWUF YH LúWDK DoÕFÕ

(45)

RODUDNNXOODQÕOÕUODU116_{Achillea türleri eucalyptol, camphor ve}_{.-terpineoller içerirler.}

.XOODQÕODQ.LP\DVDO0DGGHOHU

2,3-dimetil anilin, 2,6-dimetil anilin, 2,6-dietil anilin, 2-etil anilin, 3-etil anilin, 4-etil anilin, 2-kloro, 4-nitro anilin, 4-kloro, 2-nitro anilin, 2-anilin etanol, 4-hidroksi metil anilin, izoamil nitrit Fluka’dan, aseton Riedell’den, etanol Merck’ten, dietil eter, floro borik asit Aldrich’den, agaroz, etilendiamin tetraasetik asit (EDTA_{EDNÕU ,} NORUU EDNÕU ,, NORUU VRG\XP L\RGU DVNRUELN DVLW 6LJPD¶GDQ WLFDUL RODUDN WHPLQ edildi.

.XOODQÕODQ$OHWOHU

Jel görüntüleme sistemi (Bio Rad Gel Doc XR), elektroforez (Bio Rad), güç ND\QD÷Õ :HDOWHF 105 %UXNHU 0+] )7–IR (Mattson 1000 FTIR UNICAM Spectrometer), UV Spektrofotometre (Shimadzu, UV/Visible Recording spektrofotometre), santrifüj (Centruin 8000 Series), vorteks (Heidolph), sterilizatör +HUDHXVRWRNODYoDONDOD\ÕFÕ0HPPHUWWHUD]LS+PHWUH0HWWOHU7ROHGo), vakumlu evaporatör (RE 100B, Bibby Strilin Ltd.), membran filtresi (Schleicher&Schuell), EOHQGHUGHULQGRQGXUXFX6DQ\RPLNURGDOJDIÕUÕQYHEX]GRODEÕ$UoHOLNNXOODQÕOGÕ

3. 2. Metot

3. 2. 1. Agaroz Jel Elektroforezi

Moleküllerin sahip olduklaUÕQHWHOHNWULN\NOHULEXPROHNOOHULQELUHOHNWULNVHO DODQLoLQGHNLKDUHNHWOHULQLHWNLOHU(OHNWURIRUH]WHNQL÷LGHEXSUHQVLEHGD\DQÕU1NOHLN DVLWIUDJPHQWOHULQLQWDQÕPODQPDVÕVDIODúWÕUÕOPDVÕYHD\UÕOPDVÕLoLQNXOODQÕODQHQ\D\JÕQ yöntem agaroz jel eOHNWURIRUH]LGLU%XQHGHQOHoHúLWOLDPDoODULoLQL]ROHHGLOHQ'1$YH 51$¶ODUÕQWDQÕPODQDELOPHVLKDQJLIRUPGDROGX÷XQXQEHOLUOHQHELOPHVLE\NO÷QQ VDSWDQDELOPHVL YH |]HOOLNOH JHQHWLN PKHQGLVOL÷L WHNQLNOHUL LOH '1$ \DSÕVÕQGD ROXúWXUXODQ GH÷LúLNOLNOHUGHQ VRQUD HOGH HGLOHQ \HQL IRUPODUÕQ LQFHOHQPHVL \|QQGHQ

(46)

DJDUR]MHOHOHNWURIRUH]LWHNQL÷LPROHNOHUJHQHWLNDODQÕQGD|QHPOLELUGHQH\VHOVLVWHP ROXúWXUPDNWDGÕU

Agaroz jelde örnekler yatay pozisyonda, sabit güç ve yöndeki elektriksel alanda yürütülmektedir '1$ úHNHU IRVIDW RPXUJDVÕQGDQ GROD\Õ QHJDWLI \NOGU YH ELU HOHNWULNVHO DODQD NRQXOGX÷X ]DPDQ EX QHJDWLI \NQGHQ GROD\Õ DQRWWDQ NDWRGD GR÷UX hareket eder.

$JDUR]-HO¶LQ+D]ÕUODQPDVÕ

Agaroz (1 g) 100 mL Tris asetat tamponuna (40 mM tris asetat, 1mM EDTA, S+ LODYH HGLOGL YH PLNURGDOJD IÕUÕQGD ND\QDWÕOGÕ o_{& µ\H VR÷XWXOGX O}

HWLG\XPEURPUPJP/LODYHHGLOGLYHNDUÕúWÕUÕOGÕd|]HOWLNHQDUODUÕRWRNODYEDQGÕ LOH VDUÕOPÕú YH WDUDN \HUOHúWLULOPLú FDP WDEDND\D G|NOG -HO GRQPDVÕ LoLQ RGD VÕFDNOÕ÷ÕQGD-45 dakika bekletildi.

$JDUR]-HO(OHNWURIRUH]L<DSÕOPDVÕ -HONXOODQÕPDKD]ÕUKDOHJHOGLNWHQVRQUDRWRNODYEDQGÕDoÕOGÕYHWDUDNoÕNDUÕOGÕ +D]ÕUODQDQ'1$NDUÕúÕPODUÕX\JXQELUSLSHWLOHNX\XFXNODUDDNWDUÕOGÕ-HO00 mL Tris DVHWDWWRPSRQXP0WULVDVHWDWP0('7$S+LoHUHQHOHNWURIRUH]WDQNÕQD \HUOHúWLULOGL7DQNÕQNDSD÷ÕNDSDWÕOGÕYHHOHNWULNED÷ODQWÕODUÕ\DSÕOGÕ(OHNWURIRUH] 9¶WDP$DNÕPX\JXODQDUDNGDNLNDVUH\OH\UWOG(OHNWULNDNÕPÕNesildi, tel ED÷ODQWÕODUÕYHNDSDNoÕNDUÕOGÕ'DKDVRQUDMHOLQIRWR÷UDIÕoHNLOGL 3OD]PLG6DIODúWÕUPD

DNA kesiminin incelemenin bir yolu VXSHUFRLOHG '1$¶QÕQ VSHU NÕYUÕPOÕ oHPEHUVHO'1$NÕUÕN\RN)RUP,RSHQFLUFXODUWHN]LQFLUNÕUÕ÷ÕLoeren çembersel '1$'1$]LQFLUOHULQGHQELULQGHNÕUÕNYDU)RUP,,YH\DOLQHDUGR÷UXVDO'1$LNL ]LQFLUGH GH ELU YH\D ELUGHQ ID]OD NÕUÕN )RUP ,,, IRUPD G|QúPQ LQFHOHPHNWLU $JDUR] -HO (OHNWURIRUH]LQGH EX IRUPODU IDUNOÕ KÕ]ODUOD KDUHNHW HGHUOHU )RUP I, yük \R÷XQOX÷XID]OD KDFPL GH GúNROGX÷XLoLQ MHOGH HQ KÕ]OÕ KDUHNHW HGHU)RUP,,¶QLQ

(47)

\R÷XQOX÷XGDKDD]ROGX÷XLoLQGDKD\DYDúKDUHNHWHGHU)RUP,,,LVH)RUP,YH)RUP ,,¶QLQDUDVÕQGDELUKÕ]DVDKLSWLUùHNLO

dDOÕúPDGD NXOODQÕODQ S%OXHVFULSW 0 SOD]PLG '1$¶VÕ NE 7(16 \|QWHPLYH4LDJHQHSOD]PLGNLWLNXOODQÕODUDNVDIODúWÕUÕOGÕ

Form I Form II Form III

ùHNLO Plazmid DNA

3. 2. 2. 1. TENS Yöntemi

37 0C’ de Ampisilinli Louria Bertani (2.5g Tripton,1.25 g yeast extract, 2.5 g VRG\XP NORUU J DPSLVLOLQ P/ VDI VX VÕYÕ EHVL\HULQGH UHWLOHQ EDNWHUL NOWUQGHQ WDQH (SSHQGRUI WSH ¶HU P/ DOÕQGÕ USP¶GH GDNLND VDQWULIMOHQGLVWVÕYÕODUX]DNODúWÕUÕOGÕ+HUELUWSH/7(16çözeltisi (100 mL‘de: P/07ULV&OS+0/('7$S+J1D&OJ1D2+ eklendi ve ters düz edildi. 150 / VRG\XP DVHWDW çözeltisi (3 M pH 5.2) eklendi ve NDUÕúWÕUÕOGÕ2GDVÕFDNOÕ÷ÕQGDGDNLNDLQNEDV\RQDEÕUDNÕOGÕGDNLNDUSP¶GH santrifM\DSÕOGÕ6SHUQDWDQWWHPL]ELUWSHDNWDUÕOGÕ]HULQHP/VR÷XNPXWODNHWLO alkol ilave edildi. -200&¶GHGDNLNDLQNEDV\RQDEÕUDNÕOGÕ'DKDVRQUDUSP¶GH GDNLNDVDQWULIM\DSÕOGÕ(WLODONROX]DNODúWÕUÕOGÕSHOOHW¶OLNHWLODONROLOH\ÕNDQGÕ USP¶GHGDNLNDVDQWULIM\DSÕOGÕNWDQVRQUDDONROX]DNODúWÕUÕOGÕ(WYGH0_C’ GHDONROXQNDODQNÕVPÕX]DNODúWÕUÕOGÕYH/7(WDPSRQXLODYHHGLOGL+HUELUWüpe NRQVDQWUDV\RQJP/RODFDNúHNLOGHULERQNOHD]o|]HOWLVLLODYHHGLOGL Form II Form III Form I

(48)

3. 2. 2. 2. Mini Prep

S%OXHVFULSW 0 SOD]PLG '1$¶ VÕ 4LDJHQH SOD]PLG NLWL NXOODQÕODUDN VDIODúWÕUÕOGÕ96

S%OXHVFULSW0SOD]PLG'1$¶QÕQVDIOÕ÷ÕQDKHPHOHNWURIRUH]KHPGH 89 VSHNWURVNRSLVLQGH QP DEVRUEDQVODUÕQÕQ RUDQÕ |OoOHUHN EDNÕOGÕ '1A NRQVDQWUDV\RQXQPDEVRUEDQV|OoPLOHKHVDSODQGÕJP/Lçin A260=1.0)

'LD]RQ\XP7X]ODUÕQÕQ6HQWH]OHQPHVL

<HQLOHELOLQLUELUNOWUPDQWDUÕRODQAgaricus bisporusKLGUD]LQ\DSÕVÕQDVDKLS DJDULWLQH −Ν−[γ−L(+)-glutamil)]-4-hidroksimetilfenilhidrazin ve bunun parçalanma ürünlerini içermektedir. Agaricus bisporus’un parçalanma ürünlerinin farelerde NDQVHURMHQHWNL\HVDKLSROPDVÕYHEXSDUoDODQPDUQOHULQGHQELULQLQEHQ]HQGLD]RQ\XP L\RQXROPDVÕ87 D\UÕFD\HQLOHPH\HQELUPDQWDURODQAgaricus xanthodermus\DSÕVÕQGD p-KLGURNVLEHQ]HQGLD]RQ\XPL\RQXEXOXQPDVÕ117_{EL]LIDUNOÕVEVWLWHGLD]RQ\XPWX]ODUÕ} VHQWH]OH\LSEXQODUÕQ'1$NHVLPHWNLVLQLLQFHOHPH\H\|QHOWWL *HQHOGHGLD]RQ\XPWX]ODUÕNORUWX]ODUÕRODUDNKD]ÕUODQÕUODUIDNDWWHWUDIORURERUDW WX]ODUÕNORUWX]ODUÕQDRUDQODGDKDNDUDUOÕROGX÷XLoLQEXoDOÕúPDGDGLD]RQ\XPWX]ODUÕ tetrafloroborat tuzu olarak sentezlendi.

2,3-Dimetil benzendiazonyum tetrafloroborat (32), 2,6-Dimetil benzendiazonyum tetrafloroborat (33), 2,6-Dietil benzendiazonyum tetrafloroborat (34), 2-Etil benzendiazonyum tetrafloroborat (35), 3-Etil benzendiazonyum tetrafloroborat (36),4-Etil benzendiazonyum tetrafloroborat (37), 2-Kloro, 4-nitrobenzen diazonyum tetrafloroborat (38), 4-Kloro, nitrobenzen diazonyum tetrafloroborat (39), 2-Hidroksietil benzendiazonyum tetrafloro borat (40), 4-Hidroksimetil benzen diazonyum tetrafloroborat tuzu (41) bilinen yöntemlerle sentezlendi.89,96

(49)

.XOODQÕODQNLP\DVDOODUYHNRúXOODU øVRDP\OQLWULW+%)4 (aq.), EtOH, 0o C, 30 dakika

ùHNLO'LD]RQ\XPWX]ODUÕQÕQVHQWH]L

2,3-Dimetil benzendiazonyum tetrafloroborat (32)

2,3-Dimetil anilin (0.25 mL, 0.25 g, 2.06 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.46 mL, 0.54g, 6.19 mmol, 3.0 HTLODYHHGLOGLYHNDUÕúÕP-5 o

&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWULWP/J PPROHTGDPODGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLO eter (30 mL) ilavesiyle 2,3-GLPHWLO EHQ]HQGLD]RQ\XP WHWUDIORURERUDW NDWÕ RODUDN o|NWUOG d|NWUOHQNDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHU P/LODYHHGLOHUHNEH\D]NULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPRO% 74). e.n. 69-71 oC νmax (KBr)/cm-1 3043, 2932, 2264, 1609, 1562, 1094, 812; 1H NMR (400Mz, CD3COCD3) δ 7.12 (1H, dd, J 8.2, 1.1 Hz, ArH), 7.05 (1H, ddd, J 8.0, ,8.0, 1.1 Hz, ArH), 6.9 (1H, dd, J 8.2, 1.1 Hz, ArH), 2.28 (3H, s, CH3), 2.16 (3H, s, CH3); 13C NMR (100Mz, CD3COCD3) δ 9.7 (q), 18.6 (q), 112.2 (d), 122.9 (d), 125.2 (d), 139.1 (s), 159.9 (s), 162.3 (s) N+ -₂ BF₄ NH₂ N2 BF4 R R -+

(50)

2,6-Dimetil benzendiazonyum tetrafloroborat (33)

2,6-Dimetil anilin (0.26 mL, 0.25 g, 2.06 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.46 mL, 0.54 g, 6.19 mmol, 3.0 HTLODYHHGLOGLYHNDUÕúÕP-5 o_{&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWULWP/J}

PPROHTGDPODGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLOHWHU mL) ilavesiyle 2,6-GLPHWLO EHQ]HQGLD]RQ\XP WHWUDIORURERUDW NDWÕ RODUDN o|NWUOG d|NWUOHQNDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHU P/LODYHHGLOHUHNEH\D]NULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPROHQ 86-87 oC νmax (KBr)/cm-1 3063, 2932, 2257, 1610, 1584, 1080, 822; 1H NMR (400Mz, CD3COCD3) δ 7.06 (1H, ddd, J = 8.0, 8.0, 1.2 Hz, ArH), 6.6 (2H, dd, J = 8.3, 1.5, ArH), 2.9 (6H, s, CH3); 13C NMR (100Mz, CD3COCD3) δ 15.6 (q), 119.4 (d), 123.6 (d), 128.2 (s), 129.5 (s).

2,6-Dietil benzendiazonyum tetrafloroborat (34)

2,6-Dietil anilin (0.26 mL, 0.25 g, 2.06 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.37 mL, 0.441g, 5.03 mmol, 3.0 HTLODYHHGLOGLYHNDUÕúÕP-5 o_{&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWULWP/}

N₂ BF₄ +

-N₂ BF₄ +

(51)

-JPPROHTGDPODGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLOHWHU mL) ilavesiyle 2,6-GLHWLO EHQ]HQGLD]RQ\XP WHWUDIORURERUDW NDWÕ RODUDN o|NWUOG d|NWUOHQNDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHU P/LODYHHGLOHUHNEH\D]NULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPROHQ 99-101 oC νmax (KBr)/cm-1 3063, 2932, 2251, 1630, 1576, 1086, 816; ; 1H NMR (400Mz, CD3COCD3) δ 7.06 (1H, ddd, J = 8.0, 8.0, 1.2 Hz, ArH), 6.9 (2H, dd, J = 8.3, 1.5, ArH), 2.9 (4H, m, CH2), 2.6 (4H, m, CH3); 13C NMR (100Mz, CD3COCD3) δ 13.5 (q), 15.6 (q), 119.4 (d), 123.6 (d), 128.2 (s) 128.9 (s).

2-Etil benzendiazonyum tetrafloroborat (35)

2-Etil anilin (0.26 mL, 0.25 g, 2.06 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.46 mL, 0.54 g, 6.19 mmol, 3.0 eq) ilave HGLOGLYHNDUÕúÕP-5 o_{&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWULWP/J}

mmol, 1.2 eTGDPODGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLOHWHUP/ ilavesiyle 2-HWLO EHQ]HQGLD]RQ\XP WHWUDIORURERUDW NDWÕ RODUDN o|NWUOG d|NWUOHQ NDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHUP/LODYH HGLOHUHNDoÕNPDYLNULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPROHQ-74 o_C

νmax (KBr)/cm-1 2978, 2937, 2264, 1641, 1562, 1094, 776; 1H NMR (400Mz,

CD3COCD3) δ 9.5 (1H, dd, J 8.3, 1.5 Hz, ArH), 9.3 (1H, ddd, J 8.1, 8.1, 1.6 Hz, ArH),

9.2 (1H, dd, J 8.2, 1.1 Hz, ArH), 9.0 (1H, ddd, J 8.0, 8.0, 1.2 Hz, ArH), 3.4 (2H, m,

N₂ BF₄ +

(52)

-CH2CH3) 1.6 (3H, t, J 4.8 CH2CH3); 13C NMR (100Mz, CD3COCD3) δ 13.7 (q), 22.9

(t), 114.7 (d), 119.5 (d), 126.6 (d), 129.1 (d), 130.2 (s), 209.0 (s).

3-Etilbenzen diazonyum tetrafloroborat (36)

3-Etil anilin (0.26 mL, 0.25 g, 2.06 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.46 mL, 0.54 g, 6.19 mmol, 3.0 eq) ilave HGLOGLYHNDUÕúÕP-5 o_{&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWULWP/ g, 2.47}

PPROHTGDPODGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLOHWHUP/ ilavesiyle 3-HWLO EHQ]HQGLD]RQ\XP WHWUDIORURERUDW NDWÕ RODUDN o|NWUOG d|NWUOHQ NDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHUP/) ilave HGLOHUHNSHPEHNULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPROHQ-70 o_C

νmax (KBr)/cm-1 2970, 2932, 2270, 1642, 1470, 1080, 803; 1H NMR (400Mz,

CD3COCD3) δ 8.8 (1H, dd, J 8.3, 1.5 Hz, ArH), 8.7 (1H, ddd, J 8.1, 8.1, 1.6 Hz, ArH),

7.2 (1H, dd, J 8.2, 1.1 Hz, ArH), 6.9 (1H, ddd, J 8.0, 8.0, 1.2 Hz, ArH), 2.9 (1H, s, J 7.5, ArH) 2.6 (2H, m, CH2CH3) 1.2 (3H, t, J 4.8 CH2CH3); 13C NMR (100Mz, CD3COCD3) δ 14.0 (q) 15.1 (t), 112.4 (d), 114.6 (d), 186.9 (d), 129.2 (d), 145.7 (s), 157.3 (s). N₂ BF₄ +

(53)

-4-Etil benzendiazonyum tetrafloroborat (37)

4-Etil anilin (0.26 mL, 0.25 g, 2.06 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.46 mL, 0.54 g, 6.19 mmol, 3.0 eq) ilave HGLOGLYHNDUÕúÕP-5 o

&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWULWP/J PPROHTGDPODGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLOHWHUP/ ilavesiyle 4-HWLO EHQ]HQGLD]RQ\XP WHWUDIORURERUDW NDWÕ RODUDN o|NWUOG d|NWUOHQ NDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHUP/LODYH HGLOHUHNVDUÕNULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPROHQ-88 o C νmax (KBr)/cm-1 2978, 2932, 2264, 1641, 1584, 1086, 849; 1H NMR (400Mz, CD3COCD3) δ 7.8 (2H, dd, J 8.3, 1.5 Hz, ArH), 7.5 (2H, dd, J 8.1, 1.5 Hz, ArH), 3.0 (2H, m, CH2CH3) 1.3 (3H, t, J 4.8, CH2CH3 ); 13C NMR (100Mz, CD3COCD3) δ 13.9 (q),105.0 (t), 111.7 (s) 131. (d), 133.0 (d), 160.7 (s).

2-Kloro, 4-nitrobenzen diazonyum tetrafloroborat (38)

2-Kloro, 4-nitro anilin (0.25 g, 1.45 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.32 mL, 0.382 g, 4.35 mmol, 3.0 eq) LODYHHGLOGLYHNDUÕúÕP-5 o_{&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWULWP/J} N₂ BF₄ Cl NO₂ + -N₂ BF₄ +

(54)

-1.74 mmol, 1.2 eq) damlDGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLOHWHU mL) ilavesiyle 2-kloro, 4-QLWUREHQ]HQGLD]RQ\XPWHWUDIORURERUDWNDWÕRODUDNo|NWUOG d|NWUOHQNDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHU mL) ilave edilerek EH\D]NULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPROHQ 162-163 oC νmax (KBr)/cm-1 3063, 2991, 2276, 1641, 1562, 1080, 843; 1H NMR

(400Mz, CD3COCD3) δ 9.27 (1H, dd, J 8.3, 1.5 Hz, ArH), 9.01 (1H, dd, J 8.3, 1.5 Hz,

ArH), 8.82 (1H, s, ArH); 13C NMR (100Mz, CD3COCD3) δ 121.78 (s), 124.88 (s),

127.33 (d), 136.88 (d), 138.52 (d), 154.48 (s).

4-Kloro, 2-nitro benzendiazonyum tetrafloroborat (39)

4-Kloro, 2-nitro anilin (0.25 g, 1.45 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.32 mL, 0.382 g, 4.35 mmol, 3.0 eq) LODYHHGLOGLYHNDUÕúÕP-5 o_{&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWUDWP/J}

PPROHTGDPODGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLOHWHU mL) ilavesiyle 4-kloro, 2-QLWUREHQ]HQGLD]RQ\XPWHWUDIORURERUDWNDWÕRODUDNo|NWUOG d|NWUOHQNDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHU P/LODYHHGLOHUHNEH\D]NULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPROHQ 125-127 oC νmax (KBr)/cm-1 3063, 2967, 2284, 1649, 1562, 1344, 1173, 849; 1H NMR

(400Mz, CD3COCD3) δ 9.2 (1H, dd, J 8.3, 1.5 Hz, ArH), 8.7 (1H, dd, J 8.3, 1.5 Hz,

ArH), 7.4 (1H, s, ArH); 13C NMR (100Mz, CD3COCD3) δ 118.6 (s), 124.9 (s), 129.01

(d), 136.88 (d), 137.96 (d), 149.64 (s).

N₂ BF₄

Cl

NO₂ +

(55)

-2-Hidroksietil benzendiazonyum tetrafloro borat (40)

4-Anilin etanol (0.25 g, 1.82 mmol, 1.0 eq) etanol (1 mL) içinde çözüldü. Üzerine sulu floro borik asit (%48’lik çözelti; 0.41 mL, 0.480 g, 5.47 mmol, 3.0 eq) ilave edilGLYHNDUÕúÕP-5 o_{&µ\HVR÷XWXOGX.DUÕúÕPDLVRDPLOQLWULWP/J}

PPROHTGDPODGDPODLODYHHGLOGLYHGDNLNDNDUÕúWÕUÕOGÕ'LHWLOHWHU mL) ilavesiyle 2-KLGURNVLHWLO EHQ]HQGLD]RQ\XP WHWUDIORURERUDW NDWÕ RODUDNo|NWUOG ÇöktüUOHQNDWÕV]OHUHND]PLNWDUGDVR÷XNDVHWRQGDaP/o|]OSVR÷XNHWHU P/LODYHHGLOHUHN\HúLONULVWDOOHUKDOLQGHVDIODúWÕUÕOGÕJPPROHQ 118-120 oC νmax (KBr)/cm-1 2924, 2507, 2264, 1603, 1497, 1067, 756; 1H NMR

(400Mz, CD3COCD3) δ 7.9 (1H, dd, J 8.3, 1.5 Hz, ArH), 7.7 (1H, ddd, J 8.1, 8.1, 1.6

Hz, ArH), 7.6 (1H, dd, J 8.2, 1.1 Hz, ArH), 7.5 (1H, ddd, J 8.0, 8.0, 1.2 Hz, ArH), 3.9 (2H, t, J 4.8 CH2CH2OH) 3.48 (2H, m, J 4.8 CH2CH2OH), 1.13 (1H, t, CH2OH ); 13C NMR (100Mz, CD3COCD3) δ 54.86 (t), 55.9 (t), 57.1 (s), 60.9 (d), 123.25 (d), 130.5 (d), 134.95 (d), 141.02 (s). N₂ BF₄ OH +