Sinteza in vrednotenje zaviralcev napetostno odvisnega natrijevega kanala Nav1.3 z 2-fenil-N-(sulfamoilfenil)ciklopropanamidnim skeletom

Kastelic, Lucija (author), Zidar, Nace (mentor), Piga, Martina (comentor)

Bolečina je naravni obrambni mehanizem, ki nastane kot odziv na poškodbo ali na škodljiv dražljaj. Pri tem sodelujejo primarni senzorični nevroni v perifernem živčnem sistemu tako, da dražljaje zaznavajo preko ionskih kanalov in receptorjev. Pomembno vlogo imajo predvsem napetostno odvisni natrijevi kanali (NaV kanali), ki omogočajo nastanek in prenos bolečinskih signalov in so vključeni v več oblik kroničnih bolečinskih nevropatij. Namen magistrske naloge je bil priprava in vrednotenje novih zaviralcev napetostno odvisnega kanala NaV1.3 na osnovi koncepta hibridnih zaviralcev. Cilj je bil zasnovati molekule, ki bi se lahko učinkovito vezale na mesto za aril- in acilsulfonamidne zaviralce znotraj domene za zaznavo napetosti 4 (VSD4) in s tem potencialno dosegle visoko selektivnost in moč zaviranja NaV1.3. Pri načrtovanju smo izhajali iz že poznanih arilsulfonamidnih zaviralcev, ki so jih predhodno sintetizirali na Fakulteti za farmacijo Univerza v Ljubljani, ter iz acilsulfonamidnih zaviralcev, katerih vezavno mesto so odkrili in bolj natančno opisali pred kratkim. Sintetizirali smo šest spojin z 2-fenil-N-(sulfamoilfenil)ciklopropanamidnim skeletom. Strukturo in čistost spojin smo ovrednotili z jedrsko magnetno resonanco (NMR), masno spektrometrijo visoke ločljivosti (HRMS), infrardečo spektroskopijo (IR) in tekočinsko kromatografijo visoke ločljivosti (HPLC). Biološko vrednotenje končnih spojin so izvedli raziskovalci iz Centra za agrikulturne raziskave na Inštitutu za agrikulturo (ATK) v Mortonvasaru na Madžarskem. Uporabili so elektrofiziološko metodo vpete krpice membrane in merili zaviranje različnih podtipov NaV kanalov, izraženih v človeških embrionalnih ledvičnih celicah HEK293. Spojine so izkazale zmerno do močno zaviranje kanala NaV1.3 in zmerno selektivnost, pri čemer so nekatere pokazale prednostno vezavo na inaktivirano stanje kanala, kar je ugodna lastnost pri razvoju učinkovin za lajšanje nevropatske bolečine. Ti rezultati bodo pripomogli k načrtovanju učinkovitejših in varnejših zaviralcev NaV1.3 v prihodnosti.

• Language: Slovenian
• Keywords: bolečina, NaV1.3, napetostno odvisni natrijev kanal, načrtovanje učinkovin, sinteza, zaviralec
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FFA - Faculty of Pharmacy
• Year: 2025
• PID: 20.500.12556/RUL-176285
• COBISS.SI-ID: 258945539
• Publication date in RUL: 26.11.2025

Secondary language

• Language: English
• Title: Synthesis and evaluation of Nav1.3 voltage-gated sodium channel inhibitors with 2-phenyl-N-(sulfamoylphenyl)cyclopropanamide scaffold

Abstract

Pain is a natural defence mechanism that occurs in response to injury or harmful stimuli. In this process, primary sensory neurones of the peripheral nervous system are involved, as they detect harmful signals through ion channels and receptors. Voltage-gated sodium channels play a particularly important role, as they enable the generation and transmission of pain signals and are involved in many forms of chronic pain neuropathies. The purpose of this master’s thesis was the preparation and evaluation of new inhibitors for the voltage-gated sodium channel NaV1.3, based on the concept of hybrid inhibitors. The aim was to create molecules that could efficiently bind to the binding site for aryl- and acylsulfonamide inhibitors within the voltage-sensing domain 4 (VSD4) and potentially improve the selectivity and potency of NaV1.3 inhibition. The design of the compounds was based on previously known arylsulfonamide inhibitors synthesised at the Faculty of Pharmacy, University of Ljubljana, and acylsulfonamide inhibitors whose binding sites were recently identified and described. We synthesised six compounds with a 2-phenyl-N-(sulfamoylphenyl)cyclopropanamide scaffold. The structure and purity of the compounds were confirmed with nuclear magnetic resonance (NMR), high-resolution mass spectrometry (HRMS), infrared spectroscopy (IR) and high-performance liquid chromatography (HPLC). Biological evaluation of the final compounds was performed in collaboration with the Centre for Agricultural Research (ATK) in Mortonvasar, Hungary, using the patch clamp method to measure inhibition of different NaV isoforms expressed in human embryonal kidney cells HEK293. The final compounds exhibited moderate to strong inhibitory activity against the NaV1.3 channel, with moderate selectivity and a preference for the inactivated state, an important property in the design of drugs for the treatment of neuropathic pain. These results provide valuable information and a foundation for further development of safer and more effective inhibitors of NaV1.3.

• Keywords: pain, NaV1.3, voltage-gated sodium channel, drug design, synthesis, inhibitor