Analiza učinka niskotemperaturnih otapala na termostabilnost inzulina ; Study on the influence of deep eutectic solvents on insulin thermal stability

Strukturalna stabilnost terapeutskih proteina u tekućim formulacijama ključna je za očuvanje njihove funkcionalnosti. Novija istraživanja pokazuju da su niskotemperaturna eutektička otapala (engl. Deep Eutectic Solvents, DES) učinkovita otapala za stabilizaciji proteina. U ovom radu istražen je dizajn i svojstva DES-ova pripremljenih kombinacijom različitih donora i akceptora vodikovih veza uz varijaciju udjela vode. Analizirana su fizikalno-kemijska svojstva (gustoća, aktivnost vode, viskoznost, polarnost i pH), te računalnim metoda određeni molekulski deskriptori (σ-profili). Primjenom diferencijalne pretražne fluorimetrije utvrđeno je da određeni DES-ovi, poput kolin-klorida s glicerolom, limunskom kiselinom ili ureom te sarkozina s glicerolom, značajno povećavaju stabilnost inzulina u odnosu na referentni pufer. Dobiveni rezultati doprinose boljem razumijevanju ponašanja inzulina u DES-ovima te predstavljaju temelj za racionalni dizajn formulacija namijenjenih stabilizaciji proteina i biomolekula. ; The structural stability of therapeutic proteins in liquid formulations is crucial for preserving their functionality. Recent studies have shown that deep eutectic solvents (DES) can serve as effective media for protein stabilization. This work investigates the design and properties of DES prepared by combining different hydrogen bond donors and acceptors with varying water contents. Physicochemical properties (density, water activity, viscosity, polarity, and pH) were analyzed, and molecular descriptors (σ-profiles) were determined using computational methods. Differential scanning fluorimetry revealed that certain DES, such as choline chloride with glycerol, citric acid, or urea, as well as sarcosine with glycerol, significantly increase insulin stability compared to the reference buffer. These findings provide valuable insights into insulin behavior in DES and establish a foundation for the rational design of formulations aimed at stabilizing proteins and other biomolecules.