| Abstract | U okviru ovog doktorskog rada dizajnirani su i sintetizirani sahakini, hibridni spojevi u kojima su spojeni dijelovi
molekula antimalarika primakina, odnosno klorokina i citostatika SAHA-e (suberoilanilid hidroksamska kiselina).
Sahakini sadrže tri ključna dijela: kinolinski prsten (dio molekule primakina ili klorokina), središnji dio
(dikarboksilnu kiselinu: jantarnu, fumarnu, glutarnu, adipinsku, tereftalnu) te završni dio s dodatnom
funkcionalnom skupinom (esterskom, karboksilnom, amidnom te O-benzil-, O-metil- ili nesupstituiranom
hidroksamskom kiselinom). Sinteza sahakina polazila je od kinolinskog kraja. Primarna amino skupina primakina,
odnosno analoga klorokina i monoester odgovarajućih dikarboksilnih kiselina povezani su amidnom vezom
pomoću 1-[bis(dimetilamino)metilen]-1H-1,2,3-triazolo[4,5-b]piridinij 3-oksid heksafluorofosfata (HATU) u
prisutnosti N,N-diizopropiletilamina (DIEA) ili preko kiselinskog klorida. Dobiveni su amido-esterski derivati koji
su u sljedećem reakcijskom koraku hidrolizirani u amido-karboksilne kiseline te dalje amidirani s O-metil- ili Obenzilhidroksilaminima,
odnosno halogenanilinima (s atomima fluora/klora ili trifluorometilnom skupinom u meta
ili para položaju) uz HATU i DIEA. Sahakini sa slobodnom hidroksamskom skupinom dobiveni su katalitičkom
hidrogenolizom O-benzilhidroksamskih derivata. Svi sahakini karakterizirani su uobičajenim analitičkim i
spektroskopskim metodama (CHN, IR, 1H NMR, 13C NMR, MS). Antiproliferativno djelovanje sahakina ispitano
je na humanim tumorskim stanicama: adenokarcinoma gušterače (Capan-1), akutne mijeloične leukemije (Hap1),
karcinoma debelog crijeva (HCT-116), karcinoma pluća (NCI-H460), akutne limfoblastičke leukemije (DND-41),
akutne promijelocitne leukemije (HL-60), kronične mijeloične leukemije (K-562), multiplog mijeloma (MM.1S),
ne-Hodkinovog limfoma (Z-138), osteosarkoma (U2OS), hepatocelularnog karcinoma (HepG2), adenokarcinoma
dojke (MCF-7), karcinoma pluća ne-malih stanica (H460) i glioblastoma (U251N). Najaktivniji derivati primakina
bili su sahakini iz podskupine hidroksamskih kiselina te meta-supstituirani derivati halogenanilina s
fumardiamidnom poveznicom. Sahakinu s najjačim citostatskim učinkom, N1-hidroksi-N5-(4-((6-metoksikinolin-8-
il)amino)pentil)-glutaramidu, utvrđen je mehanizam djelovanja. Na staničnom modelu glioblastoma dokazano je da
selektivno inhibira histon-deacetilazu 6, smanjuje ekspresiju matriks-metaloproteaze-2 i receptora za epidermalni
faktor rasta. |
| Abstract (english) | This doctoral thesis describes the design and synthesis of sahaquines, hybrid molecules in which molecular parts of
antimalarial drug primaquine/chloroquine and cytostatic drug SAHA (suberoylanilide hydroxamic acid) are
merged. Sahaquines constitute of three key parts: quinoline ring (part of primaquine or chloroquine molecule),
linker (dicarboxylic acid: succinic, fumaric, glutaric, adipic, terephthalic) and the terminal part with an additional
functional group (ester, carboxylic, amide, O-benzyl-, O-methyl- or unsubstituted hydroxamic acid). Sahaquine
synthesis starts from the quinoline end. The primary amino group of primaquine/chloroquine is linked with an
appropriate dicarboxylic monoester through an amide bond achieved with 1-[bis(dimethylamino)methylene]-1H-
1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (HATU) in the presence of N,N-diisopropylethylamine
(DIEA) or by previously activating carboxylic acid with thionyl chloride. Obtained amido-esters were hydrolysed
to afford the corresponding amido-carboxylic acids, which were further coupled with O-benzyl- and Omethylhydroxylamine,
or halogenanlines. HATU/DIEA was used again as the coupling system. Free hydroxamic
acids were obtained by catalytic hydrogenation of O-benzyl derivatives. All new compounds were fully
characterized by conventional spectroscopic and analytical methods (IR, 1H and 13C NMR, MS and elemental
analyses). The cytostatic activity of sahaquines was tested on the following cancer cell lines: pancreatic
adenocarcinoma (Capan-1), chronic myeloid leukemia (Hap1), colorectal carcinoma (HCT-116), lung carcinoma
(NCI-H460), acute lymphoblastic leukemia (DND-41), acute myeloid leukemia (HL-60), chronic myeloid
leukemia (K-562), multiple myeloma (MM.1S), non-Hodgkin lymphoma (Z-138), bone osteosarcoma (U2OS),
hepatocellular carcinoma (HepG2), breast adenocarcinoma (MCF-7) and glioblastoma (U251N). The most active
primaquine derivatives were sahaquines with free hydroxamic acid moiety and diamides with meta-substituted
halogenanilines. The mechanism of action was determined for the most potent sahaquine, namely N1-hydroxy-N5-
(4-((6-methoxyquinolin-8-yl)amino)pentyl)glutaramide. On the glioblastoma cell, it is shown that sahaquine
selectively inhibits histone deacetylase 6, decreases the expression of matrix metalloproteinase-2 and the epidermal
growth factor receptor. |
