Maluck Sara, Bobrovsky Rivka, Poór Miklós, Lange Roman W, Steinmetzer Torsten, Jerzsele Ákos, Adorján András, Bajusz Dávid, Rácz Anita, Pászti-Gere Erzsébet
Department of Pharmacology and Toxicology, University of Veterinary Medicine, Hungary István utca 2, H-1078 Budapest, Hungary.
Department of Laboratory Medicine, Medical School, University of Pécs, Ifjúság útja 13, H-7624 Pécs, Hungary.
Biomedicines. 2024 Sep 11;12(9):2075. doi: 10.3390/biomedicines12092075.
Inhibitors of the serine protease furin have been widely studied as antimicrobial agents due to their ability to block the cleavage and activation of certain viral surface proteins and bacterial toxins. In this study, the antipseudomonal effects and safety profiles of the furin inhibitors MI-1851 and MI-2415 were assessed. Fluorescence quenching studies suggested no relevant binding of the compounds to human serum albumin and α-acid glycoprotein. Both inhibitors demonstrated significant antipseudomonal activity in Madin-Darby canine kidney cells, especially compound MI-1851 at very low concentrations (0.5 µM). Using non-tumorigenic porcine IPEC-J2 cells, neither of the two furin inhibitors induced cytotoxicity (CCK-8 assay) or altered significantly the intracellular (Amplex Red assay) or extracellular (DCFH-DA assay) redox status even at a concentration of 100 µM. The same assays with MI-2415 conducted on primary human hepatocytes also resulted in no changes in cell viability and oxidative stress at up to 100 µM. Microsomal and hepatocyte-based CYP3A4 activity assays showed that both inhibitors exhibited a concentration-dependent inhibition of the isoenzyme at high concentrations. In conclusion, this study indicates a good safety profile of the furin inhibitors MI-1851 and MI-2415, suggesting their applicability as antimicrobials for further in vivo investigations, despite some inhibitory effects on CYP3A4.
丝氨酸蛋白酶弗林蛋白酶的抑制剂已作为抗菌剂被广泛研究，因为它们能够阻断某些病毒表面蛋白和细菌毒素的切割与激活。在本研究中，评估了弗林蛋白酶抑制剂MI-1851和MI-2415的抗铜绿假单胞菌作用和安全性。荧光猝灭研究表明，这些化合物与人血清白蛋白和α-酸性糖蛋白无相关结合。两种抑制剂在马-达二氏犬肾细胞中均表现出显著的抗铜绿假单胞菌活性，尤其是低浓度（0.5µM）的化合物MI-1851。使用非致瘤性猪IPEC-J2细胞，即使在100µM的浓度下，两种弗林蛋白酶抑制剂均未诱导细胞毒性（CCK-8测定），也未显著改变细胞内（Amplex Red测定）或细胞外（DCFH-DA测定）的氧化还原状态。在原代人肝细胞上用MI-2415进行的相同测定，即使在高达100µM的浓度下，也未导致细胞活力和氧化应激发生变化。基于微粒体和肝细胞的CYP3A4活性测定表明，两种抑制剂在高浓度下均表现出对该同工酶的浓度依赖性抑制。总之，本研究表明弗林蛋白酶抑制剂MI-1851和MI-2415具有良好的安全性，表明它们尽管对CYP3A4有一些抑制作用，但仍适用于进一步的体内抗菌研究。
