Enhanced In Vitro Antiviral Activity of Ivermectin-Loaded Nanostructured Lipid Carriers against Porcine Epidemic Diarrhea Virus via Improved Intracellular Delivery
Xiaolin Xu, Shasha Gao, Qindan Zuo, Jiahao Gong, Xinhao Song, Yongshi Liu, Jing Xiao, Xiaofeng Zhai, Haifeng Sun, Mingzhi Zhang, Xiuge Gao, Dawei Guo
Pharmaceutics, doi:10.3390/pharmaceutics16050601
Porcine epidemic diarrhea virus (PEDV) is an acute enteric coronavirus, inducing watery diarrhea and high mortality in piglets, leading to huge economic losses in global pig industry. Ivermectin (IVM), an FDA-approved antiparasitic agent, is characterized by high efficacy and wide applicability. However, the poor bioavailability limits its application. Since the virus is parasitized inside the host cells, increasing the intracellular drug uptake can improve antiviral efficacy. Hence, we aimed to develop nanostructured lipid carriers (NLCs) to enhance the antiviral efficacy of IVM. The findings first revealed the capacity of IVM to inhibit the infectivity of PEDV by reducing viral replication with a certain direct inactivation effect. The as-prepared IVM-NLCs possessed hydrodynamic diameter of 153.5 nm with a zeta potential of -31.5 mV and high encapsulation efficiency (95.72%) and drug loading (11.17%). IVM interacted with lipids and was enveloped in lipid carriers with an amorphous state. Furthermore, its encapsulation in NLCs could enhance drug internalization. Meanwhile, IVM-NLCs inhibited PEDV proliferation by up to three orders of magnitude in terms of viral RNA copies, impeding the accumulation of reactive oxygen species and mitigating the mitochondrial dysfunction caused by PEDV infection. Moreover, IVM-NLCs markedly decreased the apoptosis rate of PEDV-induced Vero cells. Hence, IVM-NLCs showed superior inhibitory effect against PEDV compared to free IVM. Together, these results implied that NLCs is an efficient delivery system for IVM to improve its antiviral efficacy against PEDV via enhanced intracellular uptake.
to biological tests, IVM-NLCs exhibited stronger antiviral activity against PEDV than free IVM and reduced PEDV-induced mitochondrial dysfunction, which prevented ROS generation and improved viability of infected Vero cell. Moreover, IVM-NLCs also reduced PEDV-induced cell apoptosis rate. In view of the in vitro results, it would be necessary to carry out in vivo tests as soon as possible, to explore its potential in the clinical treatment of PEDV. Consequently, IVM-NLCs were demonstrated to be a potential drug against PEDV, which might provide a basis for the development of novel drugs to antagonize PEDV.
Supplementary Materials: The following supporting information can be downloaded at: https://www. mdpi.com/article/10.3390/pharmaceutics16050601/s1, Figure S1 : Cytotoxicity of Vero cells treated with different concentration of IVM at the appointed time via CCK-8 assay; Figure S2 : Antiviral activity of IVM-NLCs was measured by CCK-8 assay; Table S1 Characterization of as-prepared IVM-NLCs. Author Contributions: Conceptualization, D.G. and X.X.; methodology, X.X., S.G., Q.Z., J.G. and X.S.; formal analysis, X.X., S.G., Q.Z., J.G., X.S., Y.L., J.X., M.Z. and X.G.; investigation, X.X., S.G., Q.Z. and D.G.; resources, Y.L., J.X. and M.Z.; data curation, X.Z. and H.S.; writing-original draft preparation, X.X., S.G., Q.Z., X.S. and J.X.; writing-review and editing, D.G., J.G., Y.L., H.S., X.Z., M.Z. and X.G.; supervision, D.G., X.G., X.Z. and H.S,; project administration,..
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'abstract': '<jats:p>Porcine epidemic diarrhea virus (PEDV) is an acute enteric coronavirus, inducing '
'watery diarrhea and high mortality in piglets, leading to huge economic losses in global pig '
'industry. Ivermectin (IVM), an FDA-approved antiparasitic agent, is characterized by high '
'efficacy and wide applicability. However, the poor bioavailability limits its application. '
'Since the virus is parasitized inside the host cells, increasing the intracellular drug '
'uptake can improve antiviral efficacy. Hence, we aimed to develop nanostructured lipid '
'carriers (NLCs) to enhance the antiviral efficacy of IVM. The findings first revealed the '
'capacity of IVM to inhibit the infectivity of PEDV by reducing viral replication with a '
'certain direct inactivation effect. The as-prepared IVM-NLCs possessed hydrodynamic diameter '
'of 153.5 nm with a zeta potential of −31.5 mV and high encapsulation efficiency (95.72%) and '
'drug loading (11.17%). IVM interacted with lipids and was enveloped in lipid carriers with an '
'amorphous state. Furthermore, its encapsulation in NLCs could enhance drug internalization. '
'Meanwhile, IVM-NLCs inhibited PEDV proliferation by up to three orders of magnitude in terms '
'of viral RNA copies, impeding the accumulation of reactive oxygen species and mitigating the '
'mitochondrial dysfunction caused by PEDV infection. Moreover, IVM-NLCs markedly decreased the '
'apoptosis rate of PEDV-induced Vero cells. Hence, IVM-NLCs showed superior inhibitory effect '
'against PEDV compared to free IVM. Together, these results implied that NLCs is an efficient '
'delivery system for IVM to improve its antiviral efficacy against PEDV via enhanced '
'intracellular uptake.</jats:p>',
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