Sinteza i svojstva protuupalnih konjugata makrolida i nesteroidnih protuupalnih lijekova

doctoral thesis

• Title (english): Synthesis and activity of antiinflammatory conjugates of macrolide and nonsteroidal antiinflammatory drug
• Author: Selvira Zulfikari
• Mentor: Linda Tomašković (mentor); MBZ: 187410
• Mentor: Olga Kronja (komentor)
• Committee member: Branka Zorc (predsjednik povjerenstva)
• Committee member: Linda Tomašković (član povjerenstva); MBZ: 187410
• Committee member: Olga Kronja (član povjerenstva)
• Committee member: Sandra Jurić (član povjerenstva)
• Committee member: Ivica Malnar (član povjerenstva); MBZ: 174371
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; Zagreb
• Defense date and country: 2012-03-09, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy
• Universal decimal classification (UDC): 615 - Pharmacology. Therapeutics. Toxicology

Abstract

Nesteroidni protuupalni lijekovi (NSAID) imaju dugu kliničku primjenu u liječenju mnogih upalnih
stanja kao što su različiti oblici artritisa. NSAID-i inhibiraju sintezu prostaglandina inhibirajući enzime
ciklooksigenazu-1 (COX-1) i ciklooksigenazu-2 (COX-2). COX-1 je konstitutivno prisutan enzim odgovoran za
regulaciju normalnih fizioloških procesa, dok je COX-2 enzim fakultativno prisutan u stanjima upale, uključen
u patološkim procesima u organizmu. Međutim, ovi lijekovi pokazuju različite gastrointestinalne i
kardiovaskularne nuspojave.
Makrolidi su velika skupina različitih spojeva što se tiče njihove kemije i djelovanja, a dolaze kao
prirodni bakterijski metaboliti ili kao polusintetski derivati prirodnih spojeva. Pokazalo se da se neki makrolidi,
posebno azitromicin, nakupljaju u upalnim stanicama. Danas postoje dokazi da pojedini makrolidi, koji se već
niz godina primjenuju u kliničkoj praksi kao antibiotici, pokazuju i protuupalno djelovanje.
Cilj ovog istraživanja bio je pripraviti nove konjugate azitromicina i azitromicinu sličnih makrolida s
NSAID-ima koji pokazuju protuupalni učinak kako bi se unaprijedilo liječenje različitih upalnih bolesti.
Konjugati su pripravljeni koristeći dvostruku aktivaciju u postupku priprave esterske ili amidne veze. Korišteni
su neselektivni NSAID-i (klasični) poput indometacina, ibuprofena i flurbiprofena te selektivni COX-2 NSAIDi
poput celekoksiba i valdekoksiba. Također su kao makrolide podjedinice korišteni azitromicin, 9-deokso-9aaza-
9a-homoeritromicin A, eritromicin A, klaritromicin i telitromicin. NSAID-i su vezani na položajima 9a, 9
(oksima), 11, 3’-N, 4” i 4“-O- makrocikličkog prstena. Prva tri mjesta vezanja nalaze se na aglikonskom
prstenu, a sljedeća tri na šećerima, desozaminu, odnosno kladinozi. Također su pripravljeni i konjugati s
dekladinozilnim makrolidima kao i aglikonski konjugati. Sintetizirano je više od 90 konjugata NSAID-a i
makrolida.
Biološka aktivnost pripravljenih spojeva proučena je na temelju in vitro ispitivanja inhibicije enzima
COX-1 i COX-2 te inhibicije produkcije TNF-α (čimbenik nekroze tumora α). Pojedini konjugati pokazuju
selektivnu inhibiciju COX-2 enzima uz dodatnu inhibiciju produkcije TNF-α. Većina sintetiziranih konjugata
značajno je inhibirala produkciju TNF-α. Značajna inhibicija enzima COX-2 zabilježena je u konjugata u
kojima je NSAID vezan preko 9a-dušika na makrolidnu podjedinicu koja ima oba šećera ili na odgovarajući
aglikonski analog, te kod konjugata s telitromicinom gdje je NSAID vezan na položaju 11. Konjugati u kojima
je NSAID-na podjedinica vezana na položaju 9a pokazuju najbolji biološki učinak.

Abstract (english)

Nonsteroidal anti-inflammatory drugs (NSAIDs) have a long clinical use for the treatment of various
inflammatory diseases like different forms of arthritis. NSAIDs inhibit prostaglandin synthesis by inhibition of
enzymes cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). COX-1 was described as “constutive”
controlling normal homeostasis, and COX-2 as “inflammatory”, involved in pathological prosses in organism.
However, these drugs are not devoided of serious side effects affecting gastrointestinal and cardiovascular
system.
Macrolides form a large group of divergent compounds regarding their chemistry and function, wich
occur as natural bacterial metabolites, or as semisyntetic derivatives of natural compounds. Some macrolides,
especially azithromycin, have been shown to accumulate within inflammatory cells. Today, there is a clear
evidence that some macrolides, used for many years in clinical practice as antibiotics, also show antiinflammatory
activity.
The aim of this study was to prepare new conjugates of azithromycin and azithromycin-like macrolide
scaffolds with NSAIDs that possess anti-inflammatory activity in order to improve the treatment of different
inflammatory diseases. Conjugates have been prepared using double activation in the esters or amide coupling
procedures. Non-selective NSAIDs (classical) like indomethacine, ibuprofen, flurbiprofen and selective COX-2
NSAIDs like celecoxib and valdecoxib are used. For macrolide subunits, azithromycin, 9-deoxo-9a-aza-9ahomoerythromycin
A, erythromycin A, clarithromycin and telithromycin are used. NSAIDs were linked at the
positions 9a, 9 (oxime), 11, 3’-N, 4“- and 4”-O- of macrocyclic scaffold. The first three coupling sites are at the
aglycon part of the molecule, the later three on the sugar moieties desosamin and cladinose, respectively. Also,
conjugates of decladinosyl macrolides have been prepared, as well as macrolide aglycon conjugates. More than
90 conjugates of NSAID and macrolide have been synthesized.
Biological activity of the prepared compounds was studied on the basis of in vitro tests of COX-1 and
COX-2 inhibition and inhibition of TNF-α (tumor necrosis factor α) production. Some conjugates show
selectivity towards COX-2 enzyme and they additionally inbibit TNF-α production. Most of the synthesized
conjugates significantly inhibited TNF-α production. Significant inhibition of COX-2 was observed for
conjugates with NSAID linked through 9a-nitrogen of macrolide subunit having both sugars or for
corresponding aglycon analogues, as well as with telithromycin conjugate where NSAID is linked through
position 11. Conjugates having NSAID subunit linked at 9a position showed the highest biological activity.

• Keywords: NSAID; makrolid; konjugati; protuupalno djelovanje; COX enzim; TNF-α
• Keywords (english): NSAID; macrolide; conjugates; anti-inflammatory activity; COX enzyme; TNF-α
• Language: croatian
• URN:NBN: urn:nbn:hr:163:806085
• Study programme: Title: Pharmacy and biochemistry; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor znanosti (doktor znanosti)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2016-11-18 12:14:09