Sinteza 3-tiofenskih zaviralcev mitohondrijskih ionskih kanalčkov Kv1.3 kot protirakavih učinkovin

Mihelič, Lara (author), Tomašič, Tihomir (mentor), Gubič, Špela (co-mentor)

Napetostno odvisne kalijeve ionske kanale KV1.3 uvrščamo v skupino ionskih kanalov, ki se aktivirajo, ko pride do spremembe v napetosti membrane. Povečano izražanje kalijevih ionskih kanalov KV1.3 so odkrili v celični membrani in v notranji membrani mitohondrijev nekaterih vrst rakavih celic, kar pomeni, da lahko z razvojem zaviralcev teh kanalov v prihodnosti odkrijemo novo terapijo rakavih obolenj. V magistrski nalogi smo ponovno sintetizirali štiri znane zaviralce kalijevih ionskih kanalov KV1.3 (spojine 7, 8a, 8b in 14) in jih biološko vrednotili na rakavih in nerakavih celicah. En zaviralec (spojina 14) je v strukturi vseboval trifenilfosfonijev ion (TPP+) kot transportno skupino, ki omogoča povečano dostavo spojin v notranjost mitohondrijev, kjer ti izkazujejo zaviralno delovanje na kanalih in s tem tudi protirakavo delovanje na celicah. Trije zaviralci brez TPP+ (spojine 7, 8a in 8b) so izkazovali šibko citotoksično delovanje na celicah Panc-1 raka trebušne slinavke, medtem ko na celicah Colo357 adenokarcinoma trebušne slinavke niso imeli učinka. Spojina 14 s pripeto TPP+ je izkazovala izrazito močno citotoksično delovanje na rakavih celicah B16F10 mišjega melanoma in Colo357. Na rakavih celicah Colo357 je sprožila tudi pomembno visok nivo apoptoze. V sklopu magistrske naloge smo sintetizirali tudi dva nova zaviralca mitohondrijskih kanalov 13 in 15, ki sta v svoji strukturi vsebovala TPP+ in za en ogljikov atom daljši distančnik kot spojina 14. Zaviralca 13 in 15 sta izkazovala izrazito citotoksično delovanje na rakavih celicah B16F10 in sprožila apoptozo na rakavih celicah Colo357, pri čemer je močnejši citotoksični učinek in hitrejšo sprožitev apoptoze povzročila spojina 13. Nobena izmed ponovno sintetiziranih spojin in novih zaviralcev ni delovala specifično protirakavo, saj so spojine 7, 8a in 8b delovale citotoksično tudi na nerakavih celicah hTERT-RPE1, spojine 13, 14 in 15 pa so imele citotoksičen učinek na nerakavih celicah L929 in C2C12. Spojina 13 je sprožila apoptozo tudi na nerakavih celicah hTERT-RPE1. Spojina 13 je v svoji strukturi vsebovala cis-konfiguracijo in daljši distančnik med TPP+ in karbamatno vezjo, ki sta najverjetneje ključna elementa za močnejše protirakavo delovanje. Zaradi močnejšega citotoksičnega delovanja spojine 13 na zdravih celicah, sta se spojini 14 in 15 izkazali kot varnejša in boljša izbira za potencialno uporabo v terapiji.

• Language: Slovenian
• Keywords: Ključne besede: zaviralci kalijevih ionskih kanalov KV1.3, protirakavo delovanje, transportne skupine, tarčna dostava v mitohondrije
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2023
• PID: 20.500.12556/RUL-147244
• Publication date in RUL: 28.06.2023

Secondary language

• Language: English
• Title: Synthesis of 3-thiophene-based Kv 1.3 mitochondrial ion channels inhibitors as anticancer substances

Abstract

Voltage-gated potassium ion channels KV1.3 belong to a group of ion channels that are activated by a change in membrane potential. Increased expression of cell membrane and inner mitochondrial membrane channels is found in some specific types of cancer cells, which means that by developing inhibitors of KV1.3 channels we could potentially discover new cancer therapies. In this Master's thesis, we synthesised four known inhibitors of KV1.3 potassium channels (compounds 7, 8a, 8b and 14) and tested their anticancer activity. We also tested their cytotoxicity on normal cells. One of the compounds (compound 14) contained the triphenylphosphonium ion (TPP+) transport group, which enables better distribution of the inhibitors in the mitochondria, where they exert inhibition of the channels and anticancer activity on cancer cells. Three of the inhibitors (compounds 7, 8a and 8b) without TPP+ showed low cytotoxicity in the Panc-1 cancer cell line, while they showed no activity in Colo357 cells. Compound 14 showed strong cytotoxicity in cancer cell line B16F10 and induced a significant level of apoptosis in Colo357. The resynthesised compounds showed no specific anticancer activity. Compounds 7, 8a and 8b showed cytotoxic effects on the normal cell line hTERT-RPE1 and compound 14 showed the same effect on the normal cell lines L929 and C2C12. We also synthesised two new potential mitochondrial inhibitors 13 and 15 whose structures also contained TPP+. The compounds exhibited potent cytotoxic effects on cancer cell lines B16F10 and Colo357 and induced high levels of apoptosis in Colo357 cells. Compound 13 induced apoptosis more rapidly and its cytotoxic effect was stronger. Their anticancer effects also proved to be non-specific, exerting a strong cytotoxic effect on non-cancerous cell lines L929 and C2C12. Compound 13 also induced a high degree of apoptosis in the normal cell line hTERT-RPE1. The structure of compound 13 contained the cis-configuration and a longer linker between TPP and carbamate binding, most likely leading to a stronger anticancer activity. However, its activity in normal cell lines was also the highest among all compounds. Therefore, compounds 14 and 15 proved to be possibly better and safer choices for potential cancer therapy.

• Keywords: Keywords: inhibitors of potassium channels KV1.3, anticancer activity, transport groups