20.500.12556/DKUM-77368 Citotoksičnost in učinkovitost C87- inhibitorja tumorje - nekrotizirajočega faktorja alfa na humanih celičnih linijah diplomsko delo univerzitetnega študijskega programa I. stopnje Cytotoxicity and efficacy of C87- a tumour necrosis factor alpha inhibitor on human cell lines Bolezni, povezane s prekomernim izločanjem vnetnega citokina TNF-α, v razvitih državah predstavljajo vedno večji problem. Slaba lastnost zdravil, kot so monoklonska protitelesa (infliksimab, adalimumab in golimumab), certolizumab pegol in etanercept, ki se trenutno uporabljajo za zdravljenje teh bolezni, so različne kožne reakcije, vpliv na plodnost in potek nosečnosti, imunogenost, povečanje možnosti za nastanek kožnega raka ter visoki stroški proizvodnje. Posledično se iščejo cenejši in enako učinkoviti računalniško načrtovani mali molekularni inhibitorji TNF-α, ki so se v zadnjih letih izkazali kot potencialna rešitev tega problema. K tem računalniško načrtovanim majhnim molekularnim inhibitorjem spada tudi inhibitor C87. Inhibitor C87 so testirali na mišjih modelih s hepatitisom in ugotovili, da učinkovito inhibira TNF-α ter poveča preživetje živali. V diplomskem delu smo se odločili za preizkus inhibitorja C87 na izbrani humani celični liniji, da bi ugotovili, če obstaja dovolj velika koncentracija C87, ki ni citotoksična in ki inhibira TNF-α. Glede na rezultate smo potrdili oz. ovrgli njegov potencial kot biološko zdravilo za vnetne bolezni. Najprej smo za potrebe eksperimenta s TNF-α in C87 optimizirali eksperimentalne pogoje za uporabljene humane celične linije različnih izvorov (imunske celice, epitelne celice in celice vratu ter glave), kar pomeni, da smo posebej za adherentne in suspenzijske humane celične linije z MTT metodo določili število celic, pri katerih je bila celična viabilnost glede na koncentracijo celic najvišja. Določili smo tudi najvišje koncentracije C87, ki niso citotoksične, za vsako uporabljeno humano celično linijo posebej. V nadaljnjih eksperimentih smo se odločili uporabiti eno celično linijo, v našem primeru je to bila celična linija Jurkat, na kateri smo izvedli eksperiment s TNF-α in C87 in na podlagi katere smo izbrali koncentracijo TNF-α. Glede na rezultate eksperimenta s TNF-α in C87 smo dognali, da koncentracija 250 nM neučinkovito zavira TNF-α koncentracije 10 ng/mL. Sklenili smo, da C87 v koncentraciji 250 nM ne predstavlja potenciala za biološko zdravilo za zdravljenje boleznih, kjer je povečano delovanje TNF-α. Diseases associated with excessive secretion of the inflammatory cytokine TNF-α are a growing problem in developed countries. Disadvantages of drugs, such as monoclonal antibodies (infliximab, adalimumab and golimumab), certolizumab pegol and etanercept, currently used to treat these diseases are various skin reactions, effects on fertility and pregnancy, immunogenicity, increased risk of skin cancer and high production costs. As a result, cheaper and equally effective computer-designed small molecular inhibitors of TNF-α are being sought, which have proven to be a potential solution to this problem in recent years. These computer-designed small molecular inhibitors include the C87 inhibitor. The C87 inhibitor was tested in mouse models with hepatitis and found to be effective in inhibiting TNF-α and increasing animal survival. In this diploma, we decided to test a C87 inhibitor on selected human cell line to determine if there is a sufficiently high concentration of C87 that is not cytotoxic and that inhibits TNF-α. According to the results, we confirmed or disprove its potential as a biological cure for inflammatory diseases. First, for the needs of the experiment with TNF-α and C87, we optimized the experimental conditions for the used human cell lines of different origins (immune cells, epithelial cells and cells of the neck and head), which means that we determined the number of cells for which cell viability was relative to the cell concentration by the MTT method specifically for adherent and suspension human cell lines the highest. The highest non-cytotoxic C87 concentrations were also determined for each human cell line. In further experiments, we decided to use one cell line, in our case it was the Jurkat cell line, on which we performed an experiment with TNF-α and C87 and on the basis of which we selected the concentration of TNF-α. Based on the results of the experiment with TNF-α and C87, we found that a concentration of 250 nM ineffectively inhibited TNF-α concentrations of 10 ng/mL. We concluded that C87 at a concentration of 250 nM does not represent a potential drug for the treatment of diseases where the action of TNF-α is increased. TNF-α C87 vnetne bolezni inhibitorji TNF-α imortalizirane humane celične linije TNF-α C87 inflammatory diseases TNF-α inhibitors immortalized human cell lines true true false [A. Kralj] Slovenski jezik Angleški jezik Diplomsko delo/naloga 2020-08-27 23:19:01 2020-10-08 13:50:21 2022-08-09 15:28:37 2020-10-24 03:13:22 2020 0 Maribor 0 Maribor IX, 24 str. 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 542.978:57.083.3(043.2) 32470019 URN:SI:UM:DK:H8UUQ3A5 UN_Kralj_Adriana_2020.pdf UN_Kralj_Adriana_2020.pdf 1 80B700F0BBDCFD622758B48D12EAF416 a431e25e1b0658d5f0bc05ebf89f2d2f34695c816f716cb16c1eab38e5f97ff8 cc6ade5f-7c11-11eb-bb7a-00155d0001ca 20.500.12556/dkum/640229b4-2165-4ef1-ac4f-899579bec72b https://dk.um.si/Dokument.php?lang=slv&id=145482 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0