Sinteza novih biološki aktivnih halogeniranih hibrida bicikličkih heterocikla i 1,2,3-triazola

doctoral thesis

• Title (english): Synthesis of biologically active halogenated hybrids of bicyclic heterocycles and 1,2,3-triazole
• Author: Andrea Bistrović
• Mentor: Silvana Raić-Malić (mentor)
• Committee member: Tatjana Gazivoda Kraljević (predsjednik povjerenstva)
• Committee member: Marijana Hranjec (član povjerenstva)
• Committee member: Miroslav Bajić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2018-06-13, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry; Organic Chemistry
• Universal decimal classification (UDC): 54 - Chemistry. Crystallography. Mineralogy

Abstract

U ovom radu opisana je sinteza i biološka aktivnost novih hibrida purina i purinskih izostera s 1,4-disupstituiranim 1,2,3-triazolom. Priređeni su purini i pseudopurini sa supstituentima u položajima 6 i 9 heterocikličke jezgre i derivati purinskog izostera benzimidazola supstituiranog u položajima 2 i 5(6). 1,4-disupstituirani 1,2,3-triazoli su dobiveni Huisgenovom 1,3-dipolarnom cikloadicijom kataliziranom bakrom. Kako bi se optimirali reakcijski uvjeti, click reakcija provedena je klasičnom sintezom kao i primjenom ekološki prihvatljivih reakcija potpomognutih mikrovalovima i ultrazvukom. Svim priređenim spojevima ispitano je antitumorsko djelovanje in vitro na niz staničnih linija humanih karcinoma, kao i na zdrave stanice. Spojevi 21, 46, 56, 58, 70, 127b i 132c s najsnažnijim i selektivnim antitumorskim djelovanjem odabrani su kao kandidati za daljnja biološka ispitivanja kako bi se utvrdio mehanizam njihovog djelovanja. 5(6)-amidino-supstituirani derivati benzimidazola (125a–133a, 125b–134b, 125c–134c i 141–143) pokazali su najizraženije antitumorsko djelovanje na odabrane tumorske stanične linije u nanomolarnom području inhibitornih koncentracija. 7-deazapurinski hibridi 21, 56, 58 i 70 pokazali su značajno smanjenje ekspresije CDK9/ciklin T1 u tumorskim stanicama, dok su benzimidazolni derivati 46, 127b i 132c snažno i selektivno inhibirali karcinom pluća ne-malih stanica (A549) uz smanjenu ekspresiju enzima p38 MAPK. Daljnja in silico analiza mogućih interakcija s biološkom metom p38 MAPK potvrdila je mogućnost stvaranja značajnih interakcija s neaktivnom konformacijom tog enzima, a predviđena aktivnost imala je vrijednost nižu od 100 nM. Prema rezultatima molekulskog uklapanja priređeni su novi 5(6)-(2-imidazolinil)-supstituirani hibridi benzimidazola (153–155, 161–163, 165, 171a–175a, 171b–175b i 171c–175c) s ciljem dobivanja spojeva s boljim farmakokinetičkim svojstvima i inhibicijskom aktivnošću enzima p38 MAPK. Amidino-supstituiranim hibridima benzimidazola (125a–133a, 125b–133b i 125c–133c) ispitane su interakcije s dvolančanim polinukleotidima ctDNA i dsRNA (pApU i pCpG) primjenom UV/Vis i CD-spektroskopije, te određivanjem temperature mekšanja. Spojevima koji su pokazali mogućnost stvaranja nekovalentnih interakcija s odabranim polinukleotidima ispitano je antibakterijsko i antiprotozoalno djelovanje. Spojevi 125a i 133b pokazali su snažno antibakterijsko djelovanje protiv rezistentnih kliničkih sojeva E.coli (ESBL) i MRSA, dok je p-metoksifenil-1,2,3-triazolni supstituent (130a–130c) značajno poboljšao inhibitorno djelovanje protiv parazita T. brucei.

Abstract (english)

This thesis describes the synthesis and biological activity of novel hybrids of purine and purine isosters containing 1,4-disubstituted 1,2,3-triazoles. Purines and pseudopurines substituted at positions 6 and 9 of the heterocyclic core, as well as purine isosters benzimidazoles substituted at positions 2 and 5(6) were prepared. 1,4-Disubstituted
1,2,3-triazoles were obtained by copper-catalyzed Huisgen 1,3-dipolar cycloaddition. In order to optimize the reaction conditions, click reactions were conducted using classical synthetic approaches as well as eco-friendly microwave and ultrasound assisted reactions.
All prepared compounds were tested for their in vitro antitumor activities on several human cancer cells and normal fibroblasts. Compounds 21, 46, 56, 58, 70, 127b and 131c with the most potent and selective antitumor activities were selected as candidates for further biological studies to determine the mechanism of their action. 5(6)-Amidino substituted benzimidazole derivatives (125a–133a, 125b–134b, 125c–134c and 141–143) showed the most pronounced antitumor activities in nanomolar range towards all selected cancer cell lines. 7-Deazapurine hybrids 21, 56, 58 and 70 showed a significant reduction of CDK9/cyclin T1 expression, while benzimidazole derivatives 46, 127b and 131c strongly and selectively inhibited non-small cell lung cancer (A549) reducing p38 MAPK expression. Further in silico analysis of possible interactions confirmed the ability of significant interactions with inactive kofnormation of p38 MAPK with predicted activity below 100 nM. According to molecular docking results, new 5(6)-(2-imidazolinyl)-substituted benzimidazole hybrids (153–155, 161–163, 165, 171a–175a, 171b–175b and 171c–175c) were prepared to obtain compounds with improved pharmacokinetic properties and p38 MAPK inhibitor activities. The interactions with double-stranded polynucleotides ctDNA and dsRNA (pApU and pCpG) of chosen amidino substituted benzimidazole hybrids (125a–133a, 125b–133b and 125c–133c) were studied by using UV/Vis and CD spectroscopy and thermal denaturation experiments. Compounds that demonstrated the ability to generate
non-covalent interactions with selected polynucleotides were tested for their antibacterial and antiprotozoal activity. Compounds 125a and 133b showed strong antibacterial activity against resistant clinical strains E. coli (ESBL) and MRSA, while the p-methoxyphenyl-1,2,3-triazole substituent (130a–130c) significantly improved the activity against T. brucei.

• Keywords: purini; pseudopurini; amidini; benzimidazoli; 1 2 3-triazoli; antitumorsko djelovanje; p38 MAPK; CDK9; ctDNA; dsRNA; rezistentne bakterije; Trypanosoma brucei
• Keywords (english): purines; pseudopurines; amidines; benzimidazoles; 1 2 3-triazoles; antitumor activity; p38 MAPK; CDK9; ctDNA; dsRNA; resistant bacteria; Trypanosoma brucei
• Language: croatian
• URN:NBN: urn:nbn:hr:149:746027
• Promotion: 2018
• Project: Number: IP-2013-11-5596; Title: Sinteza i citostatska ispitivanja biblioteke novih dušikovih heterocikla; Title: Synthesis and cytostatic evaluations of novel nitrogen heterocycles library; Acronym: SCIENcENTRY; Leader: Silvana Raić-Malić; Jurisdiction: Croatia; Funder: HRZZ; Funding stream: IP
• Study programme: Title: Chemical Engineering and Applied Chemistry - Doctoral course; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti, područje prirodnih znanosti, polje kemija (doktor/doktorica znanosti, područje prirodnih znanosti, polje kemija)
• Type of resource: Text
• Extent: 268 str. ; 30 cm
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-08-30 11:01:27