Derivati 2-fenilcimetne kisline kot inhibitorji človeških hidroksiteroid-dehidrogenaz AKR1C1 in AKR1C3 : diplomska naloga

Česen, Marjeta (author), Gobec, Stanislav (mentor), Lanišnik-Rižner, Tea (comentor)

Hidroksisteroid-dehidrogenaze AKR1C1–AKR1C3 sodijo v naddruţino aldo/keto-reduktaz in so udeleţene v metabolizmu steroidnih hormonov, nevrosteroidov, ksenobiotikov in prostaglandinov. AKR1C3 katalizira redukcijo androstendiona v testosteron in estrona v estradiol, kar posredno lahko privede do povišane proliferacije celic v prostati in dojkah. AKR1C3 katalizira tudi redukcijo prostaglandina PGH2 v PGH2α in PGD2 v 9α,11β-PGF2 ter tako prepreči aktivacijo PPARγ, ki celice usmerja v apoptozo. AKR1C1 je 20-ketosteroid-reduktaza, ki pretvarja aktivni progesteron v manj aktivni 20α-hidroksiprogesteron in močan nevrosteroid 5α-pregnan-3α-ol-20-on v 5α-pregnan-3α,20α-diol. Na ta način AKR1C1 zmanjša zaščitne učinke progesterona v maternici in ektopičnem endometriju in je tako udeleţen pri nastanku raka endometrija, endometrioze in predmenstrualnega sindroma. Zaradi vpliva na metabolizem nevrosteroidov encim AKR1C1 povezujejo tudi z nastankom depresivnih motenj. AKR1C2 katalizira inaktivacijo 5α-dihidrotestosterona v 3α-androstandiol in s tem posledično ščiti celice prostate pred prekomerno proliferacijo. AKR1C1 in AKR1C3 imata pomembne vloge pri različnih patofizioloških stanjih in sta zato pomembni tarči za zdravljenje hormonsko odvisnih in hormonsko neodvisnih rakavih obolenj ter drugih bolezni. Predhodne študije so pokazale, da derivati cimetne kisline in NSAID analogi inhibirajo AKR1C izoencime. Zato smo v tej študiji preverili inhibitorno delovanje 56 strukturno sorodnih spojin iz treh različnih skupin: 1) derivate fumarne kisline, 2) derivate 3-aminobenzojske kisline in 3) derivate 2-fenilcimetne kisline na rekombinantnih encimih AKR1C1, AKR1C2 in AKR1C3. Najprej smo določili % inhibicije reakcije oksidacije 1-acenaftola, ki jo katalizirajo encimi AKR1C1, AKR1C2 in AKR1C3, v prisotnosti 100 μM posamezne spojine in 30 μM, 60 μM oziroma 100 μM substrata. Najboljši inhibitorji so bili derivati 2-fenilcimetne kisline. Spojinam, ki so encime inhibirale za več kot 50 %, smo določili vrednosti IC50. 17 spojin je imelo vrednosti IC50 v nizkem μM območju. Najboljša inhibitorja sta bili spojini 13c z 6,6 μM IC50 vrednostjo za AKR1C1 in spojina 26c z 5,8 μM IC50 vrednostjo za AKR1C3. Našli smo tudi 7 sprecifičnih inhibitorjev AKR1C3, to so bile spojine 26c, 21c, 24c, 10c, 22c, 23c in 1c z vrednostmi IC50 5,8 μM; 10,8 μM; 16,8 μM; 38,7 μM; 43,1 μM; 69,8 μM in 121,1 μM. Te spojine predstavljajo dobro izhodišče za razvoj novih specifičnih inhibitorjev AKR1C3 pri zdravljenju raka prostate in dojk ter akutne mieloidne levkemije.

• Language: Slovenian
• Keywords: hormonski sistemi hidroksisteroid-dehidrogenaze biosinteza AKR1C1 AKR1C3
• Work type: Undergraduate thesis
• Typology: 2.11 - Undergraduate Thesis
• Organization: FFA - Faculty of Pharmacy
• Place of publishing: Ljubljana
• Publisher: [M. Česen]
• Year: 2011
• Number of pages: 69 f.
• PID: 20.500.12556/RUL-121158
• UDC: 542+543
• COBISS.SI-ID: 3088497
• Publication date in RUL: 30.09.2020

Secondary language

• Language: English
• Title: Derivatives of 2-phenylcinnamic acid as inhibitors of human hydroxysteroid dehydrogenases AKR1C1 and AKR1C3

Abstract

Hydroxysteroid dehydrogenases AKR1C1–AKR1C3, members of the aldo-keto reductase superfamily, are involved in metabolism of steroid hormones, neurosteroids, xenobiotics and prostaglandins. AKR1C3 catalyzes reduction of androstenedione to testosterone and estrone to estradiol, and may thus promote the excessive proliferation of prostate and breast cells. AKR1C3 also catalyzes reduction of PGH2 to PGF2α and PGD2 to 9α,11β-PGF2 and thus prevents activation of PPARγ and cell apoptosis. AKR1C1 as 20-ketosteroid reductase inactivates progesterone by producing 20α-hydroxyprogesterone and also converts a potent neurosteroid 5α-pregnane-3α-ol-20-one to 5α-pregnane-3α,20α-diol. In this manner AKR1C1 prevents protective effects of progesterone in uterus and ectopic endometrium and is involved in promotion of endometrial cancer, endometriosis and premenstrual syndrome. Due to its role in neurosteroid metabolism it is also associated with depressive disorders. AKR1C2 catalyzes inactivation of 5α-DHT to 3α-androstanediol and thus protects prostate cells from excessive proliferation. AKR1C1 and AKR1C3 have important roles in different pathophysiological conditions so they represent important drug targets in hormone-dependent and hormone-independent cancers and other diseases. Previous studies showed that cinnamic acid derivatives and NSAID analogs inhibit AKR1C isoenzymes. Therefore, the aim of this study was to evaluate 56 compounds with similar structural characteristics from three different groups of compounds: 1) derivatives of fumaric acid, 2) derivatives of 3-aminobenzoic acid and 3) derivatives of 2-phenylcinnamic acid for their inhibition of the recombinant AKR1C1, AKR1C2 and AKR1C3 enzymes. First, we determined the percentages of inhibition of 1-acenaphthenol oxidation in the presence of 100 μM of individual compound and 30 μM, 60 μM and 100 μM concentration of substrate, respectively. The best inhibitors were derivatives of 2-phenylcinnamic acid. For compounds that showed at least 50 % inhibition of AKR1C1-AKR1C3 the IC50 values were determined. 17 compounds showed low μM IC50 values. The best inhibitor of AKR1C1 and AKR1C3 were compounds 13c and 26c with 6.6 μM and 5.8 μM IC50 values, respectively. We found 7 AKR1C3-specific inhibitors, compounds 26c, 21c, 24c, 10c, 22c, 23c and 1c, showing 5.8 μM; 10.8 μM; 16.8 μM; 38,7 μM; 43.1 μM; 69.8 μM and 121.1 μM IC50 values. These compounds represent good starting points for development of new AKR1C3-specific agents for treatment of acute myeloid leukemia and prostate and breast cancer.